KR20130113526A - Method for measuring resistance or sensitivity to docetaxel - Google Patents

Abstract

The present invention relates to novel and useful methods of predicting or monitoring a patient's response to a taxoid family of molecules by measuring the increase or decrease of certain genetic markers relative to the control. The invention also provides kits for measuring or monitoring the nucleic acid or protein levels of certain genetic markers and comparing these levels with control or reference markers to predict or monitor the patient's response to the taxoid family of molecules.

Description

Method for measuring resistance or sensitivity to docetaxel {Method for measuring resistance or sensitivity to docetaxel}

The present invention relates to a novel, useful, and hitherto unknown method for predicting or monitoring a patient's response to a taxoid family of molecules by measuring an increase or decrease in a specific genetic marker compared to a control group. The present invention also provides a kit for predicting or monitoring a patient's response to a taxoid family of molecules by measuring the nucleic acid or protein levels of a particular genetic marker and comparing these levels with a control or reference marker.

Docetaxel is an anti-mitotic agent that is widely used for the treatment of breast, lung and ovarian cancer, and to a lesser extent in the treatment of head and neck cancer, gastric cancer and prostate cancer (Hong, Oncology 16:9, 2002). Docetaxel inhibits microtubule kinetics by binding to beta-tubulin and inhibiting tumor growth by blocking the degradation of alpha- and beta-tubulin heterodimers (Ringel and Horwitz, J Natl. Cancer Inst 83:288, 1991). The antitumor activity of docetaxel and related taxanes and paclitaxel stems from targeting microtubules of the mitotic spindle, preventing the spindle from chromosome alignment and separation, blocking cell cycle progression, and activating apoptosis pathways (Wang Et al. , Cancer 88:2619, 2000). The proapoptoic activity of paclitaxel was determined by phosphorylation and disinfection of Bcl-2 through cell signaling events involving p53/p21waf1/Cip1, raf/ras, and mitogen-activated protein kinases (MAPKs). It was associated with activation (Wang et al. , Cancer 88:2619, 2000). Docetaxel is a more potent anticancer agent than paclitaxel, but the pathway involved in its cytotoxicity is not well defined (Katsumata, Br J Cancer 89:S9, 2003). Docetaxel-induced apoptosis was observed in selected cell lines through mechanisms involving Bcl-2 phosphorylation and caspase-3 activation (Kolfschoten et al ., Biochem). Pharmacol 63:733, 2002). Other proteins that modulate taxane-induced cell death in various cultured cell lines include Aurora-A (Anand et al. , Cancer) in HeLa cells. Cell 3:51, 2003), HER-2 in breast cancer cells (Tanabe et al. , Int J Oncol 22:875, 2003), p21waf1/Cip1 in glioblastoma cells (Li et al. , J Biol Chem 277:11352, 2002) and JNK/MKK1 in ovarian cancer cells (Lee et al. , J Biol Chem 273:28253, 1998).

In the present technical field, it is possible to induce the use of reagents in drug development to identify reagents (small molecules, siRNA, etc.) that inhibit the in vivo function of reagents and potentially sensitize cells to antitumor drug chemotherapy. In addition, there remains a need to identify pharmacological proteins that mediate docetaxel resistance. In addition, there remains a need in the art for an assay that can reliably predict which patients can and cannot respond to anti-tumor docetaxel drug-based chemotherapy prior to chemotherapy treatment. Applicants herein describe a novel assay that can predict resistance and susceptibility to docetaxel, and can provide a screening assay for developing a novel treatment for blocking drug resistance in cancer.

Summary of the invention

According to the present invention, a novel and useful method is provided for monitoring and/or predicting a patient's response to a taxoid family of molecules by comparing the level of activation and/or expression of specific genetic markers in patients and controls.

In one embodiment, the present invention provides a method comprising: a) obtaining a test sample from a cancerous area of a cancer patient; b) obtaining a control sample; c) measuring the level of one or more genetic markers; d) comparing the measured levels of the one or more genetic markers in the test sample and the control sample, wherein the decrease in the level of the one or more genetic markers measured in the test sample compared to the control sample is of the taxoid family. A method for predicting or monitoring a cancer patient's response to a taxoid family of molecules that exhibits increased resistance to the molecule is provided.

Specific genetic markers provided in this aspect of the present invention include the following markers:

BubR1, homo sapiens similar to protein kinase (BUBR1) mRNA, complete cds (GenBank accession number: AF046079);

Mad2, Homo sapiens mRNA for MAD2 protein (GenBank accession number: AJ000186);

Mps1, Homo sapiens TTK protein kinase (TTK), mRNA (GenBank accession number: NM_003318);

GEFT for Rac1/CDC42, Homo sapiens RAC/CDC42 exchange factor (GEFT), transcriptional variant 2, mRNA (GenBank accession number: NM_133483);

Bub1, homo sapiens BUB1 budding, mRNA (GenBank accession number: NM_004336) not inhibited by benzimidazole 1 homologue (yeast) (BUB1);

hSepharase, Homo sapiens extra spindle pole like 1 ( S. cerevisiae ) (ESPL1), mRNA (GenBank accession number: NM_012291);

CamKIId, Homo sapiens calcium/calmodulin-dependent protein kinase (CaM kinase) II delta (CAMK2D), transcriptional variant 3, mRNA (GenBank accession number: NM_001221);

CDK6, Homo sapiens cyclin-dependent kinase 6 (CDK6), mRNA (GenBank accession number: NM_001259); And

GRB2, Homo sapiens growth factor receptor-bound protein 2 (GRB2), transcriptional variant 1, mRNA (GenBank accession number: NM_002086).

In another embodiment, the present invention provides a method comprising: a) obtaining a test sample from a cancerous area of a cancer patient; b) obtaining a control sample; c) measuring the level of one or more genetic markers; d) comparing the measured levels of the one or more genetic markers in the test sample and the control sample, wherein the decrease in the level of the one or more genetic markers measured in the test sample compared to the control sample is of the taxoid family. It relates to a method for predicting or monitoring a cancer patient's response to a taxoid family of molecules that exhibits increased susceptibility to the molecule.

In this particular aspect of the invention, certain genetic markers include:

P21 (Waf1), Homo sapiens cyclin-dependent kinase inhibitor 1A (p21, Cip1) (CDKN1A), transcriptional variant 1, mRNA (GenBank accession number: NM_000389);

Pim-1, Homo sapiens pim-1 oncogene (PIM1), mRNA (GenBank accession number: NM_002648);

GBP-1, Homo sapiens guanylate binding protein 1, interferon-inducible, 67 kDa (GBP1), mRNA (GenBank accession number: NM_002053);

RXRA, Homo sapiens retinoid X receptor, alpha (RXRA), mRNA (GenBank accession number: NM_002957);

SPF45, Homo sapiens RNA binding motif protein 17 (RBM17), mRNA (GenBank accession number: NM_032905);

Hec1, kinetochore associated 2 (KNTC2), mRNA (GenBank accession number: NM_006101);

Raf1, human mRNA for raf oncogene (GenBank accession number: X03484);

Aurora A, Homo sapiens aurora-related kinase 1 (ARK1) mRNA, complete cds (GenBank accession number: AF008551);

TACC3, Homo sapiens deformable, acidic coiled-coil containing protein 3 (TACC3), mRNA (GenBank accession number: NM_006342);

RelB, Homo sapiens v-rel retinoendothelial viral oncogene homologue B, nuclear factor (algae) of kappa light chain polypeptide gene enhancer (enhancer) in B-cell 3 (algae), mRNA (GenBank accession number: NM_006509) ;

PRKCD, Homo sapiens protein kinase C, delta (PRKCD), transcriptional variant 1, mRNA (GenBank accession number: NM_006254);

BRAF35, Homo sapiens high-mobility group 20B (HMG20B), mRNA (GenBank accession number: NM_006339);

HSPA1L, Homo sapiens heat shock 70kDa protein 1A (HSPA1A), mRNA (GenBank accession number: NM_005345);

STK11, Homo sapiens serine/threonine kinase 11 (Peutz-Jeghers syndrome) (STK11), mRNA (GenBank accession number: NM_000455); And

MKK3, Homo sapiens MAP kinase kinase 3 (MKK3) mRNA, complete cds (GenBank accession number: L36719).

In a further embodiment, the present invention

a) obtaining a test sample from the cancerous area of the cancer patient,

b) BubR1, homo sapiens similar to protein kinase (BUBR1) mRNA, complete cds (GenBank accession number: AF046079);

Mad2, Homo sapiens mRNA for MAD2 protein (GenBank accession number: AJ000186);

Mps1, Homo sapiens TTK protein kinase (TTK), mRNA (GenBank accession number: NM_003318);

GEFT for Rac1/CDC42, Homo sapiens RAC/CDC42 exchange factor (GEFT), transcriptional variant 2, mRNA (GenBank accession number: NM_133483);

Bub1, homo sapiens BUB1 germination, mRNA (GenBank accession number: NM_004336) not inhibited by benzimidazole 1 homologue (yeast) (BUB1);

hSepharase, Homo sapiens extra spindle pole like 1 (S. cerevisiae) (ESPL1), mRNA (GenBank accession number: NM_012291);

CamKIId, Homo sapiens calcium/calmodulin-dependent protein kinase (CaM kinase) II delta (CAMK2D), transcriptional variant 3, mRNA (GenBank accession number: NM_001221);

CDK6, Homo sapiens cyclin-dependent kinase 6 (CDK6), mRNA (GenBank accession number: NM_001259); And

Measuring the level of one or more gene markers selected from the group consisting of GRB2, Homo sapiens growth factor receptor-bound protein 2 (GRB2), transcriptional variant 1, and mRNA (GenBank accession number: NM_002086),

c) GAPDH, Homo sapiens glyceraldehyde-3-phosphate dehydrogenase (GAPD), mRNA (GenBank accession number: NM_002046); And

RPS9, Homo sapiens cDNA clone IMAGE:6647283, measuring the level of at least one reference gene marker selected from the group consisting of partial cds (GenBank accession number: BC071941) and

d) comparing the measured levels of the one or more genetic markers and the one or more reference genetic markers in a test sample, wherein the level of the one or more genetic markers is compared to the level of the one or more reference genetic markers. It relates to a method for predicting or monitoring a cancer patient's response to a taxoid family molecule, wherein the decrease indicates increased resistance to the taxoid family molecule.

Further embodiments of the present invention

a) obtaining a test sample from the cancerous area of the cancer patient;

b) P21 (Waf1), Homo sapiens cyclin-dependent kinase inhibitor 1A (p21, Cip1) (CDKN1A), transcriptional variant 1, mRNA (GenBank accession number: NM_000389);

Pim-1, Homo sapiens pim-1 oncogene (PIM1), mRNA (GenBank accession number: NM_002648);

GBP-1, Homo sapiens guanylate binding protein 1, interferon-inducible, 67 kDa (GBP1), mRNA (GenBank accession number: NM_002053);

RXRA, Homo sapiens retinoid X receptor, alpha (RXRA), mRNA (GenBank accession number: NM_002957);

SPF45, Homo sapiens RNA binding motif protein 17 (RBM17), mRNA (GenBank accession number: NM_032905);

Hec1, Homo sapiens centromere associated 2 (KNTC2), mRNA (GenBank accession number: NM_006101);

Raf1, human mRNA for raf oncogene (GenBank accession number: X03484);

Aurora A, Homo sapiens aurora-related kinase 1 (ARK1) mRNA, complete cds (GenBank accession number: AF008551);

TACC3, Homo sapiens deformable, acidic coiled-coil containing protein 3 (TACC3), mRNA (GenBank accession number: NM_006342);

RelB, Homo sapiens v-rel retinoendothelial viral oncogene homologue B, nuclear factor of the kappa light chain polypeptide gene enhancer in B-cell 3 (algae) (RELB), mRNA (GenBank accession number: NM_006509);

PRKCD, Homo sapiens protein kinase C, delta (PRKCD), transcriptional variant 1, mRNA (GenBank accession number: NM_006254);

BRAF35, Homo sapiens high-mobility group 20B (HMG20B), mRNA (GenBank accession number: NM_006339);

HSPA1L, Homo sapiens heat shock 70kDa protein 1A (HSPA1A), mRNA (GenBank accession number: NM_005345);

STK11, Homo sapiens serine/threonine kinase 11 (Poitz-Zeros syndrome) (STK11), mRNA (GenBank accession number: NM_000455); And

Measuring the level of at least one genetic marker selected from the group consisting of MKK3, Homo sapiens MAP kinase kinase 3 (MKK3) mRNA, complete cds (GenBank accession number: L36719),

c) GAPDH, Homo sapiens glyceraldehyde-3-phosphate dehydrogenase (GAPD), mRNA (GenBank accession number: NM_002046); And

RPS9, Homo sapiens cDNA clone IMAGE:6647283, measuring the level of at least one reference gene marker selected from the group consisting of partial cds (GenBank accession number: BC071941) and

d) comparing the measured levels of the one or more genetic markers and the one or more reference genetic markers in a test sample, wherein the level of the one or more genetic markers is compared to the level of the one or more reference genetic markers. It relates to a method for predicting or monitoring a cancer patient's response to a taxoid family molecule, wherein the decrease indicates an increased sensitivity to the taxoid family molecule.

The present invention relates to a method for predicting or monitoring the response of a cancer patient to a taxoid family of molecules. Further embodiments of this invention include the taxoid family molecules paclitaxel, docetaxel XRP9881 and XRP6258.

The genetic markers described herein can be evaluated by measuring the level of RNA, DNA or protein using various techniques described in more detail below. Another embodiment of the present invention relates to a kit for predicting or monitoring a patient's response to a taxoid family of molecules.

The above and other aspects, features, and advantages of the present invention will be better understood from the following detailed description in conjunction with the accompanying drawings.

Figure 1 shows the characterization of the gene conferring its down-regulation docetaxel resistance. Docetaxel generated a dose response curve shown for HCT116 cells transfected with RB1, BubR1, Mad2 and Mps1 siRNA (■) compared to control siRNA (□). RB1 was an example of an siRNA that did not show results on the screen. Cell viability was measured at an absorbance of 450 nM after the addition of WST-1. The minimum ratio (MR) is the WST-1 value for the gene compared to the control in 40 nM docetaxel.
Figure 2 shows the dose response curve of BubR1 and Mps1 shRNA (■) compared to the control shRNA (□).
3 shows TaqMan real-time PCR analysis of BubR1 and Mps1 mRNA levels in cells containing BubR1 and Mps1 shRNA, respectively.
Figure 4 shows the cell viability and morphology shown for HCT116 cells transfected with Mad2, BubR1 and Mps1 siRNA in the presence or absence of docetaxel treatment. 24 hours after transfection, cells were left untreated (-docetaxel), or treated with 200 nM docetaxel (+docetaxel), stained with calcein-AM to visualize viable cells 16 and 72 hours after treatment. Made it.
Figure 5 shows the mitosis index for HCT116 cells transfected with Mad2, BubR1, Mps1 and control siRNA after docetaxel treatment. 24 hours after transfection, cells were treated with 200 nM docetaxel for 16 and 72 hours and tested. Mitotic cells were detected by phosphorylated histone H3 antibody, which appears as a red highlighted staining of the cell's nucleus (Mitotic Index Kit, Cellomics). The nuclei were stained blue with Hoechst dye.
6 shows TaqMan real-time PCR analysis of Mps1, BubR1 and Mad2 mRNA levels in HCT116 cells transfected with Mps1, BubR1 and Mad2 siRNA, respectively.
7 shows cell cycle analysis of HCT116 cells transfected with three mitotic checkpoint genes siRNA. After transfection with Mps1, BubR1 and Mad2 siRNA, cells were left untreated (-docetaxel) or treated with 200 nM docetaxel (+docetaxel) for 24 hours. 72 hours after addition of docetaxel, cells were harvested and analyzed for cell cycle. The number of peaks suggests DNA content such as 2N, 4N, 8N, 16N or 32N.
Figure 8 shows a clonogenic (clonogenic) cell survival analysis using a stable knockdown (knockdown) cell line. HCT116 cells containing BubR1 or vector control shRNAs were plated on 10 cm dishes and maintained for 10 days in 5 nM docetaxel. Colonies were washed with PBS and stained with crystal violet.
9 shows several dose response curves of genes whose down-regulation confers increased susceptibility to docetaxel. Docetaxel generated a dose response curve shown for HCT116 cells transfected with RB1, Pim-1, p21, Aurora A and TACC3 siRNA (■) compared to control siRNA (□). The minimum ratio (MR) is the ratio of the WST-1 reading of the gene compared to the control at 40 nM docetaxel concentration. The IC50 (IR) ratio is the ratio of the IC50 of the gene compared to the control.
10 shows the dose response curve of Aurora A shRNA (■) compared to the vector control group (□).
11 shows TaqMan real-time PCR analysis of Aurora A mRNA levels in cells containing Aurora A shRNA.
Figure 12 shows the difference in the proliferation of HCT116 cells transfected with Pim-1 and TACC3 siRNA (■) compared to the control siRNA (□). Number of cells remaining in 6-well plate at different time points after transfection with Pim-1, TACC3 and control siRNA.
13 shows TaqMan analysis of Pim-1 and TACC3 mRNA levels after siRNA transfection.
14 shows the levels of active caspase-3 in HCT116 cells transfected with Pim-1 and BubR1 siRNA. Active caspase-3 levels were expressed as fluorescence intensity using the active caspase-3 beadmates kit (Upstate). Three docetaxel concentrations of 0, 5 and 40 nM were examined at three time points 24, 48 and 72 hours after docetaxel addition.
Figure 15 shows the level of AKT phosphorylation in HCT116 cells transfected with Pim-1 and BubR1 siRNA. The ratio of phosphorylated AKT level and total AKT level was determined using a bead-based assay (Biosource). 24 hours after transfection, cells were left untreated or treated with 5 and 40 nM docetaxel. 48 hours after docetaxel treatment, the lysate was analyzed. The phosphorylation level was calculated as the ratio of phosphorylated AKT (p-AKT) to total AKT (t-AKT).
Figure 16 shows Western blot showing reduced levels of Pim-1 and BubR1 48 hours after transfection.

The present invention is broadly based on the identification by applicants of a genetic marker capable of predicting whether a test subject is resistant or susceptible to a drug of the Taxoid family. Using a cell-based RNA interference (RNAi) screen, Applicants observed that drug resistance or susceptibility to docetaxel was associated with certain genetic markers.

It should be noted that a number of terms and phrases have been used throughout the specification and claims. Definitions of these terms and phrases are given below:

As used herein, the term “prognosis” refers to drug resistance, and the prediction or probability of cancer-causing death or progression, including relapse, metastatic spread and neoplastic disease.

As used herein, the term “forecast” refers to the likelihood that a patient may respond favorably or undesirably to a drug or set of drugs, and the patient's surgical removal of the primary tumor and/or cancer recurrence after chemotherapy. It refers to the degree of reaction that is such as to be able to survive for a certain period of time without this. The predictive method contemplated by the present invention can be used clinically to determine the treatment regimen by selecting the most appropriate treatment modality for a particular patient, particularly taxoid-based chemotherapeutic agents. The predictive method of the present invention is whether the patient will respond favorably to a treatment regimen such as chemotherapy and/or radiotherapy with a given drug or drug combination, or after the end of chemotherapy or other treatment modalities. It is a useful means of predicting whether or not a person will survive.

As used herein, the term “tumor” refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all precancerous and cancerous cells and tissues.

As used herein, the terms “cancer” and “cancerous” generally refer to a physiological condition in a mammal characterized by unregulated cell growth. Examples of cancer include, but are not limited to, the following cancers: breast cancer; Colon cancer; Lung cancer; Prostate cancer; Hepatocellular carcinoma; Stomach cancer; Pancreatic cancer; Cervical and ovarian cancer; Liver cancer; Bladder cancer; Urethral cancer; carcinoma; Melanoma; Brain cancer, including glioblastoma and glioblastoma; Biliary tract cancer; Chorionic cancer; Esophageal cancer; Stomach cancer; Hematologic neoplasms including acute lymphoid and myeloid leukemia; Multiple myeloma; AIDS-associated leukemia and adult T-cell leukemia lymphoma; Intraepithelial neoplasms including Bowen's disease and Paget's disease; Lymphoma including Hodgkin's disease and lymphocytic lymphoma; Neuroblastoma; Oral cancer including squamous cell carcinoma, leiomyosarcoma, rhabdomyosarcoma, liposarcoma, fibrosarcoma, and sarcoma including osteosarcoma; Skin cancers including melanoma, Kaposi's sarcoma, basocellular cancer and squamous cell carcinoma; Testicular cancer, including embryonic tumors such as endothelial tumors, non-epithelial tumors (teratoma, chorionic carcinoma), interstitial tumors, and germ cell tumors; Thyroid cancer, including thyroid adenocarcinoma and weeping cancer; And renal cancer, including adenocarcinoma and Wilms tumor.

As used herein, the term “patient” preferably means human, but may also include non-human primates, cattle, horses, pigs, sheep, goats, dogs, cats or rodents. Preferably, the subject is suspected of having cancer, is diagnosed with cancer, or is a high-risk group for the onset of cancer, eg, a human with a family history of cancer. In a preferred embodiment of the invention, the cancer is breast cancer. Methods of identifying a subject suspected of having cancer include manual examination, tissue biopsy, family history of the subject, medical history or mammography of the subject, magnetic resonance imaging, magnetic resonance spectroscopy, or positron emission tomography. A number of imaging techniques, such as, may be included. Methods for diagnosing cancer and the clinical characterization of cancer diagnosis are well known to experts skilled in the medical field.

As used herein, the term "sample" is a tissue obtained using a method well known to an expert of ordinary skill in the relevant medical technology field. Methods such as tissue biopsy include mass, microdissection, laser-based microdissection, or the total distribution of other cell-separating methods known in the art. Due to the variability in cell type and sensitivity of the diagnostic method used in the tissue biopsy material of diseased tissue, the sample size required for analysis is 1, 10, 50, 100, 200, 300, 500, 1000, 5000, It may range from 10,000, to 50,000 or more cells. Appropriate sample size can be determined based on the conditions of the cell composition and tissue biopsy, and the standard preparation steps for this determination and subsequent isolation of nucleic acids for use in the present invention are well known to those skilled in the art. It is known. For example, a sample from a tissue biopsy may be sufficient to assess RNA expression without amplification. Conversely, in the absence of a suitable number of cells in a small tissue biopsy area, it may be necessary to use RNA conversion and/or amplification methods or other methods that promote degradation of nucleic acid molecules. Such methods, which allow the use of limited tissue biopsy materials, are well known to those of ordinary skill in the art. Some examples include, but are not limited to, direct RNA amplification, reverse transcription of RNA to cDNA, amplification of cDNA, or generation of radio-labeled nucleic acids.

As used herein, the term “test” in relation to a sample refers to a sample taken from a cancerous area of the body, or from an area of the body that exhibits a stage or characteristic of cancer.

As used herein, the term "control" related to a sample means a sample used for comparison purposes. Preferably, these samples are "in the sense that the samples do not exhibit symptoms of any disease or condition that may affect gene expression, or are believed to have these diseases or conditions, particularly with respect to the diseases for which they will be used as standards. It is a control". Instead, it can be understood that different stages of a disease or condition may be compared, in which case the “control” sample is one that corresponds to an earlier stage of the disease or condition. For example, the control sample can be a sample taken from an equivalent area of the body that is non-cancerous. Additionally, the control sample may be an equivalent area of the body from the same subject, or a non-cancerous area of the body from a second subject substantially similar to the first subject (same or similar species, age, weight, sex, etc.). Finally, a control sample can also be taken from a cancerous area of a second subject that responds well to treatment with a taxoid family of molecules.

A "nucleic acid molecule" refers to a ribonucleoside (adenosine, guanosine, uridine or cytidine "RNA molecule") or a deoxyribonucleoside (deoxyadenosine, deoxyguano), which is a single stranded form or a double-stranded helix. Phosphate ester polymeric form of a new, deoxythymidine or deoxycytidine; "DNA molecule"), or any phosphoester homologues thereof such as phosphorothioates and thioesters. There may be double stranded DNA-DNA, DNA-RNA and RNA-RNA helices. The term nucleic acid molecule, and in particular a DNA or RNA molecule, refers only to the primary and secondary structure of the molecule and is not limited to any particular tertiary form. Thus, the term includes, inter alia, linear or circular DNA molecules (eg restriction fragments), plasmids and double-stranded DNA present in chromosomes. When referring to the structure of a particular double-stranded DNA molecule, the sequence is only a standard that presents the sequence in the 5'to 3'direction along the non-transcriptional strand of DNA (i.e., the strand having a sequence homologous to the mRNA). It may be described herein in accordance with the Convention. A "recombinant DNA molecule" is a DNA molecule that has undergone molecular biological manipulation.

As used herein, the term "portion" of an isolated nucleic acid molecule encoding a particular protein refers to a portion or fragment of an isolated nucleic acid molecule comprising a sufficient number of contiguous nucleotides encoding a peptide or polypeptide. do. Naturally, a "portion" of an isolated nucleic acid molecule is more than one nucleotide, and the peptide or polypeptide encoded by this portion contains a number of amino acid residues as described in the definitions of peptides and polypeptides below. .

As used herein, the term "peptide" refers to two or more amino acids covalently linked by a peptide bond. In certain embodiments, the peptide comprises at least 10, preferably at least 20, more preferably at least 30, even more preferably at least 40, most preferably 50 or more amino acids.

As used herein, the term "polypeptide" refers to a linear polymer composed of a number of contiguous amino acids. In particular, the polypeptide may have a molecular weight greater than 100 kD.

As used herein, the term "genetic marker" refers to a physiological composition that functions to predict whether the level of RNA, DNA or protein thereof measured in a sample is resistant or susceptible to a drug of the Taxoid family. Means. Moreover, a genetic marker may encode a particular protein, or may instead act as a “surrogate” marker for a protein whose activity is related to the level of a genetic marker in a sample of body origin. This relationship can be direct, as the case where a decrease in the level of protein activity corresponds to a decrease in the level of a genetic marker, or instead this relationship is that a decrease in the level of protein activity corresponds to an increase in the level of the genetic marker. Correspondingly, it could be the opposite relationship. Such physiological compositions include, but are certainly not limited to, cellular (eg, progenitor stem cell) proteins, polypeptides, DNA, RNA, carbohydrates, or fatty acids to mention just a few. In certain embodiments of the present invention, the measured level of a certain genetic marker can predict whether a test subject is resistant or susceptible to a drug of the taxoid family. Examples of such genetic markers include, but are certainly not limited to the following markers:

BubR1, homo sapiens similar to protein kinase (BUBR1) mRNA, complete cds (GenBank accession number: AF046079);

Mad2, Homo sapiens mRNA for MAD2 protein (GenBank accession number: AJ000186);

Mps1, Homo sapiens TTK protein kinase (TTK), mRNA (GenBank accession number: NM_003318);

GEFT for Rac1/CDC42, Homo sapiens RAC/CDC42 exchange factor (GEFT), transcriptional variant 2, mRNA (GenBank accession number: NM_133483);

Bub1, homo sapiens BUB1 budding, mRNA (GenBank accession number: NM_004336) not inhibited by benzimidazole 1 homologue (yeast) (BUB1);

hSepharase, Homo sapiens extra spindle pole like 1 (S. cerevisiae) (ESPL1), mRNA (GenBank accession number: NM_012291);

CamKIId, Homo sapiens calcium/calmodulin-dependent protein kinase (CaM kinase) II delta (CAMK2D), transcriptional variant 3, mRNA (GenBank accession number: NM_001221);

CDK6, Homo sapiens cyclin-dependent kinase 6 (CDK6), mRNA (GenBank accession number: NM_001259); And

GRB2, Homo sapiens growth factor receptor-bound protein 2 (GRB2), transcriptional variant 1, mRNA (GenBank accession number: NM_002086);

P21 (Waf1), Homo sapiens cyclin-dependent kinase inhibitor 1A (p21, Cip1) (CDKN1A), transcriptional variant 1, mRNA (GenBank accession number: NM_000389);

Pim-1, Homo sapiens pim-1 oncogene (PIM1), mRNA (GenBank accession number: NM_002648);

GBP-1, Homo sapiens guanylate binding protein 1, interferon-inducible, 67 kDa (GBP1), mRNA (GenBank accession number: NM_002053);

RXRA, Homo sapiens retinoid X receptor, alpha (RXRA), mRNA (GenBank accession number: NM_002957);

SPF45, Homo sapiens RNA binding motif protein 17 (RBM17), mRNA (GenBank accession number: NM_032905);

Hec1, Homo sapiens centromere associated 2 (KNTC2), mRNA (GenBank accession number: NM_006101);

Raf1, human mRNA for raf oncogene (GenBank accession number: X03484);

Aurora A, Homo sapiens aurora-related kinase 1 (ARK1) mRNA, complete cds (GenBank accession number: AF008551);

TACC3, Homo sapiens deformable, acidic coiled-coil containing protein 3 (TACC3), mRNA (GenBank accession number: NM_006342);

RelB, Homo sapiens v-rel retinoendothelial viral oncogene homologue B, nuclear factor (algae) of kappa light chain polypeptide gene enhancer (enhancer) in B-cell 3 (algae), mRNA (GenBank accession number: NM_006509) ;

PRKCD, Homo sapiens protein kinase C, delta (PRKCD), transcriptional variant 1, mRNA (GenBank accession number: NM_006254);

BRAF35, Homo sapiens high-mobility group 20B (HMG20B), mRNA (GenBank accession number: NM_006339);

HSPA1L, Homo sapiens heat shock 70kDa protein 1A (HSPA1A), mRNA (GenBank accession number: NM_005345);

STK11, Homo sapiens serine/threonine kinase 11 (Peutz-Jeghers syndrome) (STK11), mRNA (GenBank accession number: NM_000455); And

MKK3, Homo sapiens MAP kinase kinase 3 (MKK3) mRNA, complete cds (GenBank accession number: L36719).

As used herein, the term "reference genetic marker" refers to a physiological composition in which its measured RNA, DNA or protein levels remain unchanged before, during or after exposure to a taxoid family of drugs. “Reference genetic markers” are called housekeeping genes. These are genes selected on the basis of relatively constant levels of expression in the system tested, for example in certain diseases such as cancer. Housekeeping genes are used to standardize the results of expression. Examples of such genetic markers include, but are certainly not limited to the following markers:

GAPDH, Homo sapiens glyceraldehyde-3-phosphate dehydrogenase (GAPD), mRNA (GenBank accession number: NM_002046); And

RPS9, Homo sapiens cDNA clone IMAGE:6647283, partial cds (GenBank accession number: BC071941).

As used herein, the term "taxoid family of molecules" refers to a class of chemotherapeutic compounds belonging to the taxane family. Specific members of the taxoid family include paclitaxel (taxol), docetaxel (taxotere) and their homologues (i.e., XRP9881 and XRP6258; see: Ojima and Geney, Curr. Opin Investig Drugs 4:737, 2004). Because these classes of molecules are beta-tubulin binding agents and stabilize the polymerized form of microtubules similar to docetaxel, the clinical expression of the biomarkers described here may reflect similar reaction states to these drugs. It is expected to be able to.

As used herein, the term “molecule”, “compound” or “pharmaceutical” refers to any composition currently known or found later. Examples of compounds or drugs having applicability in the present invention include organic compounds (e.g., synthetic, naturally occurring, optically active compounds), peptides (synthetic, naturally occurring, optically active, That is, D or L amino acids), carbohydrates, nucleic acid molecules, and the like.

As used herein, "stringency of hybridization" or "hybridization under stringent conditions" means a condition that can be easily determined by those of ordinary skill in the art, and is generally These are empirical calculations depending on the probe length, wash temperature, and salt concentration. In general, longer probes require higher temperatures for proper annealing, while shorter probes require lower temperatures. Hybridization generally depends on the ability of denatured DNA to reanneal when complementary strands are present in an environment below their melting point. The greater the degree of desired homology between the probe and the hybridizable sequence, the higher the relative temperature that can be used. As a result, higher relative temperatures tend to make the reaction conditions more stringent, while lower temperatures make them less stringent. For further details and description of the stringency of hybridization reactions, see Ausubel et al., Current Protocols in Molecular Biology, Wiley Interscience Publishers, (1995).

As used herein, “stringent conditions” or “high stringency conditions” refer to (1) low ionic strength and high temperature for washing, eg, 0.015 M sodium chloride/0.0015 M sodium citrate/0.1 at 50° C. % Sodium dodecylsulfate or (2) denaturing agents, e.g. formamide, e.g. 50% (v/v) formamide and 0.1% bovine serum albumin/0.1% ficoll, at 42° C. during hybridization (Ficoll)/0.1% polyvinylpyrrolidone/pH 6.5 50 mM sodium phosphate buffer and 750 mM sodium chloride, 75 mM sodium citrate, or (3) 50% formamide at 42° C., 5x SSC (0.75 M NaCl, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5x Denhardt's solution, sonicated salmon semen DNA (50 .mu.g/ml), 0.1 % SDS, and overnight hybridization in solution using 10% dextran sulfate, washing for 10 minutes in 0.2x SSC (sodium chloride/sodium citrate) at 42° C., then 0.1x SSC with EDTA at 55° C. It refers to a parameter for washing with a 10-minute high-stringency washing solution consisting of. Appropriate stringent conditions can be identified as described in Sambrook et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Press, 1989, and less stringent washing solutions and hybridization conditions than those described above ( Examples include the use of temperature, ionic strength and% SDS). Examples of suitably stringent conditions are 20% formamide, 5x SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5x Denhardt's solution, 10% dextran sulfate and 20 mg/ml. Incubate overnight at 37[deg.] C. in a solution containing denatured sheared salmon semen DNA, and then the filter is washed in 1x SSC at about 37-50[deg.] C.. The skilled practitioner will recognize how to adjust temperature, ionic strength, etc. as needed to adjust factors such as probe length and the like. The stringency suitable for hybridizing a nucleic acid depends on the length of the nucleic acid and the degree of complementarity, parameters well known in the art. The greater the degree of similarity or homology between two nucleotide sequences, the greater the value of Tm for hybridization of nucleic acids having these sequences. The relative stability of nucleic acid hybridization (corresponding to a larger Tm) decreases in the following order: RNA:RNA, DNA:RNA, DNA:DNA. Preferably, the minimum length for hybridizable nucleic acids is at least about 12 nucleotides, preferably at least about 16 nucleotides, more preferably at least about 24 nucleotides in length, and most preferably at least 36 nucleotides. .

As used herein, "label" or "detectable-label" is directly or indirectly conjugated to an antibody, oligopeptide, or other organic molecule to form a "labeled" antibody, oligopeptide or other organic molecule. It means a detectable labeling compound or composition to be produced. The label may be detectable by itself (eg, a radioisotope label or a fluorescent label), or, in the case of an enzymatic label, may facilitate chemical modification of the detectable substrate compound or composition.

As used herein, "direct label" refers to a material that can be easily visualized with the naked eye in its natural state, or with the aid of an optical filter and/or applied stimulus, for example, ultraviolet light that promotes fluorescence. . Examples thereof include, but are not limited to, colored labels, metallic sol particles, dye sol particles dyed latex, or dye-encapsulated liposomes (U.S. Patent No. 4,313,734, U.S. Patent No. 4,373,932, WO 88/ 08534, EP-A 0 280 559, 0 281 327, described in U.S. Patent No. 4,703,017). Other direct labels include radionucleotides, radiopaque substances, fluorescent or luminescent sites.

As used herein, "indirect label" also refers to an enzyme that can be used according to the present invention. Various types of enzyme-linked immunological assays are well known in the art, for example alkaline phosphatase and horseradish peroxidase, lysozyme, glucose-6-phosphate dehydrogenase, lactate dehydrogenase. Genases, ureases, these and others are described in detail in Eva Engvall, Enzyme Immunoassay ELISA and EMIT in Methods in Enzymology, 70:419-439 (1980) and U.S. Patent No. 4,857,453.

As used herein, the term “kit” refers to an article comprising one or more containers and a label or package insert on or associated with the container. In a preferred embodiment, the container may contain a detectable-labeled antibody, a detectable-labeled antibody fragment or a detectable-labeled oligonucleotide. In another embodiment, the container obtains total RNA from a sample of body origin, reverse transcription of the total RNA to obtain cDNA, and the cDNA is a polymerase using a set of detectably-labeled primers. It provides a means of applying to chain reactions. For example, additional containers of kits containing diluents and buffers, control antibodies, oligopeptides or small organic molecules may be included. Labels or product instructions may provide a description of the composition and instructions for storage and intended in vitro or diagnostic use.

In addition, according to the present invention, conventional molecular biology, microbiology and recombinant DNA techniques within the scope of the art can be used. These techniques are described in detail in the literature (see e.g. Sambrook, Fritsch & Maniatis, Molecular Cloning: A Laboratory. Manual , Second Edition (1989) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York (hereinafter "Sambrook et al., 1989"); DNA Cloning : A Practical Approach , Volumes I and II (DN Glover editors. 1985); Oligonucleotide Synthesis (Editor, MJ Gait. 1984); Nucleic Acid Hybridization [Editors BD Hames & SJ. (1985)]; Transcription And Translation [BD Hames & SJ Higgins, editors. (1984)]; Animal Cell Culture [RI Freshney, editor. (1986)]; Immobilized Cells And Enzymes [IRL Press, (1986)]; B. A Practical Guide To Molecular Cloning (1984); FM Ausubel et al. (eds.), Current Protocols in Molecular Biology , John Wiley & Sons, Inc. (1994)).

In general, RNAi technology uses synthetic or vector-generated double-stranded RNA to induce degradation of mRNA containing homologous sequences (McManus and Sharp, Nat Rev. Genet 3:737, 2002). Using RNAi screening, Caenorhabditis elegans (Simmer et al. , PloS Biol 1:E12, 2003), Drosophila (Lum et al. , Science 299:2039, 2003) and mammalian cells (Aza-Blanc et al. , Mol. Cell 12:627, 2003).

In the present invention, HCT116 colon cancer cells were used to screen siRNAs directed against 101 cancer-related genes, including most of the members of the kinase family, and docetaxel lethal effect was determined by a high-resolution dose-response curve. Was quantified by. Using this approach, Applicants found that under-expression of nine genes including BubR1, Bub1, Mad2, Mps1 and GEFT Rac/CDC could inhibit the lethal effect by docetaxel, while It has been shown that low-expression of 15 other genes including Pim1, p21, TACC3 and Aurora-A can elevate docetaxel-induced cell death.

Accordingly, the present invention broadly extends to diagnostic methods and kits that can be used to determine whether an individual taking or considering taking docetaxel is resistant or susceptible to this drug.

Applicants have observed drug resistance to docetaxel, whereby drug resistance is associated with the loss of an expanded set of checkpoint control genes including BubR1, Bub1, Mad2, Msp1 and GEFT Rac/CDC.

Increased docetaxel sensitivity is one of several genes including GBP-1, STK11, RXRA, Hec1, SPF45, Raf1, RELB, MKK3, PRKCD, HSPA1A, BRAF35, Aurora-A, Pim-1, TACC3 and p21waf1/Cip1. Corresponding to the loss, inhibitors to these genes can serve as useful adjuvants available for docetaxel therapy.

The mitotic checkpoint gene is delayed by the inhibition of Cdc20-APC (anaphase-promoting complex) until the congeners of the sister chromatids are properly attached to the mitotic spindle and aligned to the equator. By acting as a failsafe mechanism (Zhou et al. , J Cell Sci 115:3547, 2002), allowing cells to reliably exit mitosis and enter an additional cell cycle with the correct complement of DNA. In the presence of microtubule inhibitors, cells from mitosis are no longer able to properly separate their chromosomes and can generally undergo apoptosis immediately after or after several cell cycles (Taylor and McKeon, Cell 89:727, 1997). The combination of microtubule inhibitors and loss of mitotic checkpoint genes such as BubR1 allows cells to avoid mitotic arrest without undergoing apoptosis and induce chromosomal instability (CIN) and aneuploidy (Shin Etc. , Cancer Cell 4:483, 2003). Recently, drug resistance data were generated when treating BubR1 and Mad2 down-regulated MCF-7 cells with the microtubule inhibitor paclitaxel (Sudo et al. , Cancer). Res 64:2502, 2004).

The Pim-1 gene encodes a serine/threonine kinase that belongs to a small group of related kinases. Pim-1 function is associated with proliferation, differentiation, apoptosis, tumorigenesis, hypoxia, angiogenesis and mitosis (Wang et al. , Biochim Biophys Acta 1593:45, 2002). Microscopic imaging data along with AKT hypophosphorylation data and measurement of increased caspase-3 activity indicate that Pim-1 down-regulation promoted docetaxel-induced apoptosis by inactivating AKT signaling that mediates cell survival. . Pim-1 and p21 have very similar characteristics in sensitizing HCT116 cells to docetaxel, which is consistent with reports indicating that p21 is the phosphorylating substrate of Pim-1 (Wang et al. , Biochim). Biophys Acta 1593:45, 2002). Pim-1 down-regulation induced some apoptosis in the absence of docetaxel, but was more effective in the presence of the drug, whereas down-regulation of the deformable acidic coiled-coil protein TACC3 induced cell death in the absence of docetaxel. Was not done, indicating that RNAi alone has no effect on cell proliferation. These results suggest that depletion of different proteins can induce different mechanisms to sensitize cells to docetaxel.

Thus, in one aspect of the present invention, the activation of one or more of the genes described in Tables I and II and the protein levels of these genes can be used to select a therapeutic treatment strategy and monitor the effectiveness of the selected treatment strategy.

In certain embodiments, methods are provided for assessing and predicting the effectiveness of chemotherapy in a patient by monitoring the level of activation of one or more genes shown in Tables I and II during the course of chemotherapy. DNA (HafnerEtc.,Arch Pathol Lab Med 127:1221, 2003; RodriguezEtc.,Clin Cancer Res 10:5785, 2004), messenger RNA (Chang et al., Lancet. 362:340, 2003) and/or protein (EspinaEtc.,J Immunol Methods 290:121, 2004) tissue biopsies from tumors (e.g., breast cancer, colon cancer, non-small cell lung cancer, and gastric tumors), or tumor tissue from undiagnosed cancer patients or patients currently undergoing treatment. Short cultivation of biopsies is performed to assess the level of gene activation or protein level of the genes or subsets thereof described in Tables I and II. Gene transcription and protein expression profiling have utility in predicting and diagnosing therapeutic responses to potential therapeutic strategies, and can be useful tools for monitoring a patient's response to therapy.

According to the present invention, RNA can be isolated from tumor samples using any available method commonly used in the art (Ullmann et al., J Biomol Screen 9:95, 2004; Badiee et al., BMC. Biotechnol 3:23, 2003). One or more cells are obtained from test subjects and RNA is isolated from these cells. For example, peripheral blood leukocytes (PBLs) cells may be obtained from a subject, or a cell sample may be obtained or the sample may be enriched for the desired cell type. Cells may be isolated from a cell mixture using a variety of techniques, such as cell separation using antibodies that bind to a specific epitope on the desired cell type. When the desired cells are solid tissue, certain cells can be excised, for example, by microdissection or by laser entrapped microdissection (LCM) (Bonner et al. , Science 278:1481, 1997; Fend Et al. , Am J Path 154:61, 1999). RNA can also be extracted from tissue or cell samples by various methods, for example guanidinium thiocyanate lysis followed by CsCl centrifugation (Chirgwin et al. , Biochemistry 18:5294, 1979). RNA from single cells can be obtained as described in the method for preparing cDNA libraries from single cells (Dulac, Curr Top Dev Biol. 36:245, 1998). RNA samples can be enriched for specific species. In one embodiment, for example, poly(A)+RNA can be isolated from an RNA sample. In particular, poly-T oligonucleotides can be immobilized on a solid support to act as an affinity ligand for mRNA. Kits for this purpose can be used as commercial products, for example, MessageMaker kit Life Technologies, Grand Island , NY). Enrichment can be achieved, for example, by primer-specific cDNA synthesis, or by several rounds of linear amplification and template-directed in vitro transcription based on cDNA synthesis (Wang et al. , Proc. Natl Acad . Sci USA 86:9717, 1989; Dulac et al., supra).

For example, PCR; Ligase chain reaction (LCR) (Wu and Wallace, Genomics 4:560, 1989); Self-sustained sequence replication SSR (Guatelli et al. , Proc Natl Acad Sci USA 87:1874, 1990); Nucleic acid base sequence amplification (NASBA) and transcription amplification (Kwoh et al. , Proc Natl Acad Sci USA 86:1173, 1989) are suitable for use in the method of the present invention. Methods for PCR technology are well known in the art (see, e.g., PCR Technology: Principles and Applications for DNA Amplification (ed. HA Erlich, Freeman Press, NY, NY, 1992); PCR Protocols: A Guide to Methods and Applications (eds. Innis, et al., Academic Press, San Diego, Calif., 1990)).

RNA can be labeled for detection using any method known to those skilled in the art, for example using microarrays such as those made by Affymetrix. RNA is labeled and detected by hybridization using any commonly used method, such as direct RNA labeling with dyes such as cyanine-3 or cyanine-5 (Perkin Elmer MPS544001KT). Further methods include reverse transcription using nucleotides labeled with cyanine-3 or cyanine-5 (Badiee A 2003), fluorescein or Alexa dyes (Molecular Probes), or other fluorophores. Direct cDNA labeling is included. In addition, ^{indirect cDNA labels such as FairPlay TM} / aminoallyl label (Stratagene Catalog number: 252002), 3DNA dendrimer label (Genisphere Inc) or enzymatic signal amplification (MICROMAX TSA, Perkin Elmer) are used. Can be. Alternatively, RNA can be quantified using RNA probes that are detected by enzymatic, fluorescent, radioactive, or light emission methods.

In one aspect of the invention, RNA is obtained from a tumor sample, and Taqman technology is used for detection of genetic markers. Primers are generated to detect specific RNAs specified in Tables I and II, and amplified using reverse transcriptase. Detection of specific fragments amplified by reverse transcription is achieved by gel electrophoresis, polymer electrophoresis, direct DNA sequencing, photodetection, loss of fluorescence signal, enzymatic reaction, or detection by hybridization to complementary RNA. .

In another aspect of the present invention, oligonucleotides are detected using real-time PCR. The mRNA obtained from the tumor sample is reverse-translated into cDNA. Primers are generated to amplify specific RNAs or subsets thereof specified in Tables I and II using the polymerase chain reaction technique. Detection of mRNA levels is achieved by gel electrophoresis and ethidium bromide or CYBR green staining, or by measuring the fluorescence intensity from fluorophore-labeled sequence specific probes released by polymerase.

In a further aspect of the present invention, a Northern blotting technique is used. RNA is obtained from a tumor sample, then separated by gel electrophoresis and transferred onto a membrane. RNA abundance is determined by hybridization using a radioisotope labeled probe such as P32, or by enzyme-based pigment productivity or luminescence assay.

In yet another further aspect of the invention, the gene dosage can be used as a surrogate indicator of the transcript level. In Tables I and II, the gene dose of the transcript or a subset thereof can be quantitatively measured and evaluated as to whether or not the tumor is responsive to docetaxel therapy (Rodriguez et al ., Clin). Cancer Res 10:5785, 2004). Because these assessments can be derived prior to adjuvant (preoperative treatment) therapy, patients can be treated with docetaxel if they are classified as responders, or another if they are classified as non-responders. Chemotherapeutic agents may be administered.

DNA is extracted from patient tumor tissue biopsy and purified to remove contaminants using the same method as is performed by experts skilled in the art. Methods used to determine gene dose in tumor DNA include, but are not limited to, quantitative PCR, genomic DNA-chip, in situ hybridization or Southern (Hafner et al. , Arch Pathol Lab Med 127:1221, 2003; Rodriguez et al ., Clin Cancer Res 10:5785, 2004). Thus, these methods can be used to determine the copy number of one or more genes in Tables I and II and compared to a control sample or a reference marker.

In another aspect of the invention the protein level is used as a surrogate measure of the transcript level. Protein levels have been shown to correlate with inhibition or elevation of transcript levels. Accordingly, the present invention includes techniques for measuring the level of a protein corresponding to the polypeptide product encoded by the transcripts in Tables I and II. The protein is detected and used as a marker of the predicted response to docetaxel. Methods of detecting proteins are well known in the art. For example, an immunological assay is a method of detecting the expression of a target protein using an antibody.

In certain embodiments, immunological assays are used to detect protein products of one or more genes listed in Tables I and II. Additionally, antibodies against any of the proteins listed in Tables I and II can be used in a number of other detection methods. These detection methods include, but are not limited to, the following methods: Western blots, ELISA assays, sandwich ELISA. Instead, other analytical methods that do not use antibodies are contemplated, including methods of using DNA oligonucleotides or polypeptides (e.g., aptamers) that recognize proteins encoded by transcripts in Tables I and II. This includes. Further, mass spectrometry of a biological sample may be used to determine the composition of the polypeptide by measuring the precise molecular weight of the peptide fragment after proteolytic cleavage.

The present invention further provides an enzyme, a fluorophore (e.g., fluorescein isothiocyanate (FITC), phycoerythrin (PE), Texas red (Texas red TR), rhodamine, free or chelated lanthanide). Class salts, chromophores, radioisotopes, chelating agents, dyes, colloidal gold, latex particles, ligands (eg biotin) and chemiluminescent agents.

In one aspect of the present invention, the radiolabel used may be isotopes such as 3H, 14C, 32P, 35S, 36 Cl, 51 Cr, 57Co, 58Co, 59Fe, 90Y, 125 I, 131I, and 186Re. Quantitative analysis of radioactivity levels can be performed through currently known and available counting methods. Conversely, radiopaque materials may be used. In embodiments in which the label is an enzyme, detection can be performed by any of the colorimetric, spectrophotometric, fluorescence spectrophotometric, amperometric or gas quantification techniques known in the art and currently used.

A further aspect of the invention is the use of a peptide binding agent such as an antibody or fragment of an antibody. Antibodies include polyclonal and monoclonal antibodies prepared according to conventional methods. Only a small portion of the antibody molecule, the paratope, is involved in the binding of an antibody to its epitope (generally, Clark, WR (1986) The Experimental Foundations of Modem Immunology Wiley & Sons, Inc., New York; Roitt, I. (1991) Essential Immunology, 7th Ed., Blackwell Scientific Publications, Oxford). For example, the pFc' and Fc regions are effectors of the complement cascade, but are not involved in antigen binding. An antibody designated as an F(ab')2 fragment, whose pFc' region is enzymatically cleaved or produced without a pFc' region, retains both the antigen binding sites of the intact antibody. Thus, embodiments of the present invention utilize detectably-labeled antibody fragments such as F(ab')2 fragments. Similarly, an antibody designated as a Fab fragment, whose Fc region is enzymatically cleaved or produced without an Fc region, retains one of the antigen binding sites of the intact antibody molecule. The Fab fragment consists of a portion of the antibody light chain that is covalently linked and the antibody heavy chain denoted Fd. Fd fragments are the main determinants of antibody specificity (a single Fd fragment can bind to 10 different light chains without changing antibody specificity), and the Fd fragments are isolated and retain epitope-binding capacity.

As is well known in the art, in the antigen-binding portion of an antibody, a complementary determinant region (CDR) that directly interacts with an epitope of an antigen, and a framework region (FR) that maintains the tertiary structure of the paratope. ) Exists (see generally Clark, 1986; Roitt, 1991). In both the heavy chain Fd fragment and the light chain of an IgG immunoglobulin, there are four framework regions (FR1 to FR4) separated by three complementary determining regions (CDR1 to CDR3), respectively. The CDRs, and particularly the CDR3 regions, and more particularly the heavy chain CDR3, are largely responsible for antibody specificity. It is well established in the art that non-CDR regions of mammalian antibodies can be substituted by similar regions of homologous or heterologous antibodies while retaining the epitope specificity of the original antibody. This is most evident in the development and use of “humanized” antibodies that covalently bind non-human CDRs to human FR and/or Fc/pFc' regions to produce functional antibodies (see US Patent No. 4,816,567, 5,225,539, 5,585,089, 5,693,762 and 5,859,205).

Fully human monoclonal antibodies can also be prepared by immunizing transgenic mice for most of the human immunoglobulin heavy and light chain positions. After immunization of these mice (eg XenoMouse Abgenix, HuMAb mice (Medarex/GenPharm)), monoclonal antibodies can be prepared according to standard hybridoma techniques. These monoclonal antibodies may have a human immunoglobulin amino acid sequence and thus will not cause a human anti-mouse antibody (HAMA) response when administered to humans.

Therefore, as readily apparent to those skilled in the art, the present invention also provides F(ab')2, Fab, Fv and Fd fragments Fc and/or FR and/or CDR1 and/or CDR2 and/or Chimeric antibody FR and/or CDR1 and/or CDR2 and/or light chain CDR3 region in which the light chain CDR3 region is replaced by a homologous human or non-human sequence. ab')2 fragment antibody FR and/or CDR1 and/or CDR2 and/or light chain CDR3 region is replaced by a homologous human or non-human sequence Chimeric Fab fragment antibody and FR and/or CDR1 and/or CDR2 region It includes chimeric Fd fragment antibodies that have been replaced by homologous human or non-human sequences. The present invention also includes so-called single chain antibodies.

The invention may be better understood by reference to the following non-limiting examples provided as examples of the invention. The following examples are presented to more fully describe the preferred embodiments of the present invention. However, they are not to be understood as limiting the broad scope of the invention in any way.

[Example]

Example 1: siRNA screening

To identify genes that make cells more resistant or susceptible to docetaxel treatment, siRNAs for 101 genes were screened in HCT116 cells using a range of docetaxel concentrations to generate a dose-response curve. Human colon cancer cell line HCT116 was obtained from ATCC, 95% CO2 and 5% O ₂ At 37° C. under 100 units/ml penicillin, 100 mg/ml streptomycin, 4 mM L-glutamine and 10% fetal bovine serum were incubated in McCoy 5A supplemented. The siRNA list consisted of cancer-related genes, including those detected by gene expression profiling experiments, that were overexpressed in docetaxel-resistant breast tumors (Chang et al. , Lancet 362:362, 2003). An additional criterion was whether the target was suitable for drug development. The overall plan is summarized as follows. On the first day, HCT116 cells were plated at 5,000 cells per well in a 96-well format and transfected the next day with a pool of 3-4 siRNA per gene. Representative samples of the siRNA used are shown in Tables I and II. HCT 116 cells were transfected with siRNA (Dharmacon) in 96 well plates using Lipofectamine 2000 (Invitrogen). On the third day, siRNA was removed, and docetaxel was added at a concentration ranging from 0 to 40 nM. On day 6, cell viability was quantified using the WST-1 assay. This is an assay to monitor the activity of mitochondrial dehydrogenase present in viable cells, which causes the delivery of the tetrazolium salt WST-1 to form formazan. On the day of analysis, media was removed from the 96 well plate. WST-1 reagent (Roche) was diluted 10-fold in McCoy 5A medium, and 100 μl was added to each well. The plates were then incubated at 37° C. for 40-80 minutes and then read at 450 nm on SpectroMax (Molecular Devices). The WST-1 value was collected in stages by calculating the ratio of the experimental siRNA to the control siRNA, and a total of 15 genes were specified to confer sensitivity, and 9 genes conferring resistance are shown in Table III below. The susceptible and tolerant populations were further divided into additional groups: 1) siRNA effects were observed at low concentrations (1-6 nM) of docetaxel resulting in fluctuating IC50, or 2) main effects were observed at higher concentrations (> 6 nM). When IC50 fluctuations are small.

Compared to Mad2, BubR1 and Mps1 siRNAs, siRNAs targeting Grb2, CDK6, Sepharose, and calcium/calmodulin-dependent protein kinase II delta (CamKIID) have characteristic fluctuations in IC50 and lower docetaxel concentrations (1 To 6 nM) showed a more remarkable effect. Overall, this subgroup showed weaker tolerance than the mitotic checkpoint gene, and CamKIID siRNA provided a maximum level of protection, 2-fold compared to control cells. These data demonstrate that loss of four mitotic checkpoint genes along with several other genes can increase docetaxel resistance to varying degrees. The data in Figure 1 show several characteristic dose-response curves for cells transfected with siRNA and then exposed to various concentrations of docetaxel. Four mitotic checkpoint genes including BubR1, Bub1, Mad2, and Mps1 (data not shown for Bub1) and siRNA to the guanine nucleotide exchange factor (GEFT) for Rac/Cdc42 showed significant docetaxel resistance, The effect was more pronounced at higher docetaxel concentrations (>6 nM). The maximum level of resistance was shown for Mad2, when cell viability increased 5-fold compared to control siRNA transfected cells. Most siRNAs have not been evaluated on the screen, and human retinoblastoma 1 (RB1) is shown as a representative example here.

Taken together, these results show that siRNA screening differs in how down-regulating gene expression sensitizes cells to docetaxel treatment or protects them from docetaxel treatment, and how it modulates docetaxel-mediated cell death depending on the concentration. It shows that it can provide insight into whether it is driving the mechanism.

[Table I]

[Table II]

[Table III]

Example 2: Cross validation for a subset of siRNAs conferring increased docetaxel resistance

Potential drawbacks to siRNA screening methods are that the siRNA anneals to a transcript with partial identity or initiates a micro-RNA (miRNA) translation block, which results in a target-specific effect and non-specific (off-site). It is based on observations that it can make the effects difficult to distinguish (Jackson et al. , Nat Biotechnol 21:635, 2003). To confirm resistance using an independent method using the second sequence, stable knockdown cell lines for BubR1 and Mps1 were generated by infecting HCT116 cells with a retroviral vector carrying a DNA oligonucleotide encoding a short hairpin (sh)RNA. Made it. The shRNA contained a different sequence than the siRNA by targeting a separate site in the open reading frame. Retrovirus packaging cell line GP2-293 was obtained from Clontech. Cells were maintained in DMEM supplemented with 100 units/ml penicillin, 100 mg/ml streptomycin, 4 mM L-glutamine and 10% fetal bovine serum. Viruses were generated by transiently transfecting GP2-293 cells. A total of 3.6 x 10 ⁶ cells were inoculated into 10 cm dishes 24 hours before transfection. Four hours prior to transfection, the medium was replaced with DMEM containing 10% FBS without antibiotics. Cells were transfected with 6 mg of vector DNA, 6 mg of envelope plasmid VSV-G (Clontech) and 72 ml of Lipofectamine-2000. The medium was changed after 14-16 hours; Virus supernatant was harvested after 24 hours, filtered through a 0.45 mM filter, and used to infect HCT116 cells at an MOI of 5 in the presence of 8 mg/ml polybrene. 48 hours after infection, cells were selected from 0.5 mg/ml puromycin for 7 days or until background cells died. Again, the dose response curve generated using the BubR1 and Mps1 knockdown cell lines showed more markedly enhanced tolerance at higher docetaxel concentrations, similar to the results obtained by siRNA transient transfection (FIG. 2 ). In order to test whether shRNAs reduced mRNA levels, the present inventors detected endogenous transcripts using real-time PCR, and confirmed that RNA levels were reduced in BubR1 and Mps1 stable knockdown cell lines compared to stable vector control cell lines ( Fig. 3).

Cells were lysed and RNA was extracted using RNAqueous-96 (Ambion). Taqman ^™ probes and forward and reverse primers were ^{designed using Primer Express™} software (PE Applied Biosystems, UK). BLAST exploration of probe and primer sequences did not reveal significant identity to sequences other than the specific gene under test. For real-time PCR, the following 20 ml of master TaqMan ^™ mixture was prepared for each well: 4.825 ml Rnase-free water, 12.5 ml 2x Universal PCR master mixture and 0.625 ml 40x Multiscribe ^™ and Rnase inhibitor mixture (Applied BioSystems), 0.9 ml forward and reverse primers (100 mM), 0.25 μl probe (100 mM). 5 μl of sample RNA (approximately 1 ng/ml) was added to each well. ^{Plates were analyzed on a TaqMan™} ABI Prism 7700 Sequence Detecter™ (Perkin-Elmer, UK). The cycling parameters were 48° C. for 30 minutes, 95° C. for 10 minutes, 40 cycles of 95° C. for 15 seconds, and 60° C. for 1 minute. The test gene mRNA values were extrapolated from the standard curve and expressed as the remaining %.

Example 3: Down-regulation of Mad2, BubR1 and Mps1 protects cells from docetaxel-induced death by avoiding mitotic arrest coupled with the production of aneuploids.

As measured by WST-1 assay, loss of the mitotic checkpoint gene in the presence of docetaxel was associated with increased cell viability indicating resistance (FIG. 1 ). In order to further test the effectiveness of the WST-1 measurement, cytological experiments were performed to investigate cell morphology and viability (FIG. 4). Cells were tracked using confocal microscopy and calcein-AM, a biostaining pigment that is converted to green fluorescence by intracellular esterases that act as indicators of metabolic active cells. FIG. 4 shows the mitosis of Mad2, BubR1 and, to a lesser extent, Mps1 siRNA transfected cells 16 hours after docetaxel addition, to an apparent intermittent state consistent with the early, flattened cell morphology, whereas Most of the control siRNA transfected cells indicated that they were arrested by rounded cell morphology in mitosis. The squamous cell phenotype has already been described for other microtubule inhibitors, which indicates the ability of cells to escape from mitotic arrest and into an apparent intermittent state without actually completing mitosis (Kung et al., Proc. Natl Acad Sci 87:9553, 1990; Lanni and Jacks, Mol Cell Biol 18:1055, 1998). The most surprising observation for Mad2, BubR1 or Mps1 siRNA down-regulated cells 72 hours post treatment is the presence of multiple more cells and their very large compared to siRNA control transfected cells substantially eliminated by docetaxel treatment. It was size. In the absence of docetaxel, mitotic checkpoint down-regulated cells appeared similar to control cells at two different time points, and an increased number of cells at 72 hours indicated that the cells were actively growing in the presence of siRNA. Suggested.

Mitosis checkpoint-damaged cells can bypass mitotic arrest and escape mitosis early by treatment with microtubule inhibitors such as colsemide, nocodazole and paclitaxel (Taylor and McKeon, Cell 89:727, 1997; Shin et al. , Cancer Cell 4:483, 2003; Masuda et al. , Am J Pathol 163:1109, 2003; Sudo, etc. , Cancer Res 64:2502, 2004). To determine whether docetaxel-treated HCT116 cells escape from mitosis due to inhibition of three specific checkpoint genes Mad2, BubR1 or Mps1, the present inventors measured the mitotic index of the transfected cells. Therefore, siRNA-transfected Mad2, BubR1 or Mps1 cells were harvested without treatment with docetaxel (0), or 8, 16, 24, 36, 48 and 72 hours after treatment with docetaxel, and then fixed and mitotic As an indicator, it was cultured with a polyclonal antibody that detects phosphorylation of histone H3 (Mitotic Index Kit, Cellomics). The graphical representation for all time points is shown in FIG. 5. The mitotic index was highest between 16 and 24 hours after treatment, suggesting that there was mitotic arrest in all cases. The mitotic index reached the highest value for cells transfected with control siRNA, while cells transfected with Mad2 and BubR1 siRNA showed a significant decrease in mitotic index. The Mps1 siRNA effect was not as severe as Mad2 and BubR1. These results clearly indicate that mitotic checkpoint damaged cells can bypass mitotic arrest induced by docetaxel, with BubR1 siRNA being the most effective, followed by Mad2 and Mps1.

In order to exclude the possibility that the change in the mitotic index coincided with a large difference in knockdown efficiency, the present inventors measured the mitotic checkpoint transcription level using real-time PCR. The experiment shown in FIG. 6 shows that the mRNA levels were reduced to equivalent levels in Mad2, BubR1 and Mps1 siRNA transfected cells, suggesting that the mitotic index difference was more reflective of gene function.

Microscopy data showed that very large cells can accommodate abnormal DNA content. To further investigate whether these mitotic checkpoint damaged cells can reverse docetaxel-induced cell cycle arrest, and to reenter the S-phase to generate aneuploid cells, the present inventors performed FACS analysis to determine the DNA content. Was measured. Mps1, BubR1, Mad2 and control siRNA transfected cells were grown in the absence or presence of docetaxel, fixed, and then stained with propidium iodide and analyzed by FACS. Data was collected at several different time points after addition of docetaxel (data not shown), and a histogram for the 72 hour time point is shown in FIG. 7. 72 hours after addition of docetaxel, transfection with Mps1, BubR1, Mad2 and siRNA caused accumulation of cells with 32N DNA content in the case of 8N, 16N and BubR1. By using only control siRNA, an 8N population of cells was detected in the presence of docetaxel, but the number of 8N was much greater for mitotic checkpoint down-regulated cells. At an earlier time point after docetaxel addition (data not shown), most of the control cells were arrested at 4N, while cells transfected with Mps1, BubR1 and Mad2 siRNA progressed from 4N to 8N. These results suggest that knockdown of the mitotic checkpoint gene allows cells to accumulate abnormal DNA levels faster and cross a greater threshold of ploidy compared to wild-type cells. Our results demonstrate a role for Mad2, BubR1 and Mps1 in regulating mitotic index and cell cycle progression by previously unspecified microtubule inhibitor docetaxel.

Example 4: Formation of docetaxel resistant colonies in BubR1 down-regulated cells

The apoptosis function of BubR1 is related to the regulation of chromosomal fidelity, considering that underexpression of BubR1 along with antimicrobial agents was associated with aneuploid and repair of BubR1 was associated with activated checkpoints and apoptosis. It seems to be important (Shin et al. , Cancer Cell 4:483, 2003). To determine whether permanently down-regulating the mitotic checkpoint gene can grow cells with greater proliferative capacity, BubR1 and Mps1 stable knockdown cell lines (Figure 4) were used to proliferate and settle colonies in the continuous presence of docetaxel. Let's test their abilities. Mad2 stable knockdown cell lines could not be generated because Mad2 damage was fatal to the cells (Michel et al. , Proc. Natl Acad Sci 101:4459, 2004). BubR1, Mps1 and control shRNA stable knockdown cell lines were applied to 5 nM docetaxel for 10 days and then stained with crystal violet. 8 shows that the BubR1 knockdown cell line produced a number of large colonies compared to the control cell line. The Mps1 knockdown cell line was unable to grow colonies in the presence of 5 nM docetaxel. When the plate was cultured for a long period of time, such as 17 days, colonies were observed on the control plate, but the number was much smaller compared to the BubR1 knockdown cell line. These results associate low levels of BubR1 with advanced cell growth in the presence of the chemotherapeutic drug docetaxel, but not Mps1.

Example 5: Cross validation of siRNA conferring increased docetaxel sensitivity

In addition to identifying siRNAs that protect cells from docetaxel-induced death, the screen identified siRNAs that sensitize cells to death. In Table III, Pim-1, p21 ^{waf1 / Cip1} , GBP-1, RXRA, Hec1, SPF45, Raf1 are a group of siRNAs showing marked killing at higher docetaxel concentrations (>6 nM) with _{only small fluctuations in IC 50.} Represents. The siRNA dose-response curves for the tumorigenic serine/threonine kinase Pim-1 and the cell cycle inhibitor p21 ^{waf1 / Cip1} were very similar to be able to reflect a redundant or cooperative role within the signaling pathway (FIG. 9 ); Previous reports have indicated that p21 ^{waf1 / Cip1} is a substrate for Pim-1 (Wang et al. , Biochim Biophys Acta 1593:45, 2002). Of further interest were data on Pim-1 overexpression in prostate epithelial cells leading to mitotic checkpoint interference and gene instability (Roh et al. , Cancer Res 63:8079, 2003). These results are consistent with Mad2, BubR1 and Mps1 underexpression data indicating comparable phenotypes with opposite gene expression levels (see Figure 1). Another siRNA that confers susceptibility to docetaxel was Hec1.

In Table III, TACC3, RELB, Aurora-A, PRKCD (PKC), HSPA1A, and BRAF35 show siRNAs that increase sensitivity at lower docetaxel concentrations (1-6 nM) with characteristic fluctuations in IC50. Aurora-A is a serine/threonine kinase that acts on the onset of mitosis and has been implicated in centrosome maturation and spindle assembly (MeraldiEtc.,Curr Opin Genet Dev 14:29, 2004). Of great interest, consistent with our findings that underexpression of Aurora-A enhanced docetaxel lethality, published data showing that Aurora-A overexpression in HeLa cells protected the cells from paclitaxel (AnandEtc.,Cancer Cell 3:51, 2003). Obviously, this was a small decrease in IC50 for Aurora-A indicating a 25% decrease in survival; However, the results were accurately doubled by using retroviral delivery of shRNAs resulting in stable knockdown of Aurora-A in HCT116 cells (compare FIG. 9 with FIGS. 10 and 11).

Example 6: Pim1 inhibits the proliferation of HCT116 cells in the absence of docetaxel.

To further assess positivity from the screen, it was important to distinguish between siRNAs that affect cell viability in the absence of docetaxel and those that act cooperatively with docetaxel to promote killing. To distinguish the possibility, we used a trypan blue exclusion assay with a cell count to determine the number of viable cells in a 6-well plate at 24, 48, and 72 hours after transfection to siRNA against proliferation. The effect of transfection was studied. Our results showed that Pim-1 siRNA inhibited growth in HCT116 cells, whereas TACC3 siRNA had no effect on growth (Fig. 12; knockdown quantified in Fig. 13), but both siRNAs were docetaxel. -Induced cell death was enhanced (Fig. 9).

Example 7: Verification of a subset of resistance and susceptibility genes using survival/kill assays

In order to visually compare the siRNA that confers docetaxel sensitivity and resistance, microscopic imaging is used to measure the ratio of the dead cells to the viable cells. Here, the viable cells are stained green using calcein-AM, and Sodium iodide (PI) stains the nuclei of dead cells in red. After docetaxel addition (72 hours), siRNA-transfected BubR1 cells had a larger number of viable cells, while siRNA-transfected Pim-1 and TACC3 had the largest number of killed cells, with control cells in between. (Data not shown). Although BubR1 siRNA alone increased cell death by 72 hours, the cells ultimately became more resistant to docetaxel. Pim-1 siRNA alone induced cell death up to 24 hours, whereas TACC3 siRNA did not induce cell death. These results demonstrate that BubR1 down-regulation is associated with increased cell viability and drug resistance, while Pim-1 and TACC3 down-regulation are associated with increased cell death and drug sensitivity. Both Pim-1 and TACC3 showed more killing in the presence of docetaxel, but this analysis could not differentiate between additive effects and cooperative killing.

Example 8: Docetaxel-induced caspase-3 activity was reduced by BubR1 down-regulation and enhanced by Pim-1 down-regulation.

Docetaxel can induce cell death through induction of apoptosis (Kim et al. , Int J Mol Med 11:799, 2003). To determine whether Pim-1 and BubR1 siRNA were able to modulate the canonical apoptosis pathway in the absence and presence of docetaxel, the inventors studied the activated caspase-3 levels in HCT116 cells (Bio Using bioplex assay) (Figure 14). 48 hours after 40 nM docetaxel treatment, caspase-3 activity was induced in control cells, which induction was further enhanced by down-regulating Pim-1 and reduced by down-regulating BubR1 (center and lower panels). . In the absence of docetaxel, down-regulation of Pim-1 caused a slight increase in caspase-3 activity at 24 hours, but returned to control levels by 48 and 72 hours (upper panel), which is another apoptosis. This suggests that the device was activated and maintained the enhanced killing observed at 72 hours by WST-1 and life/death analysis. Down-regulation of BubR1 in the absence of docetaxel caused a very slight increase in caspase-3 activity, which is consistent with the observation that BubR1 knockdown primarily caused cell death at 72 hours in the live/death assay. . These results further include Pim-1 and BubR1 as effectors of docetaxel killing through modulation of caspase-3 activity.

Example 9: Docetaxel-induced Pim-1-related sensitivity and signaling pathways involved in BubR1-associated resistance

The activity attributable to Pim-1 suggests that it plays a pivotal role in blocking signaling events that induce cell death while promoting promoting cell survival. In an attempt to define the molecular basis for enhanced cell death associated with Pim-1 knockdown, the present inventors examined the activation state of AKT, a pivotal cellular pathway involved in cell survival. HCT116 was inoculated on a 6-well plate at 4×10 ⁵ cells/well, and the next day, it was transfected with siRNA at 16 nM using Lipofectamine 2000 (Invitrogen). After 24 hours, cells were treated with or without 5 and 40 nM docetaxel. Ice-cold lysis buffer (50 mM HEPES buffer (PH7.4), 1% NP40, 2.5 mM EDTA, 100 mM sodium fluoride, 10 mM sodium PP _I , protease inhibitor cocktail) 24, 28 and 72 hours after docetaxel addition Purified (Roche), 2 mM sodium orthovanadate), and centrifuged at 12,000 g for 10 minutes. Thereafter, the lysate was quantitatively analyzed for the total protein level using DC protein assay (BioRad). Active caspase-3 levels were ^{measured using the Luminex 100™} system and active caspase-3 beadmates kit (Upstate) according to the protocol presented by the manufacturer. Total and phosphorylated AKT levels were ^{measured using a Luminex 100™} system and a total AKT antibody bead kit and a phosphospecific AKT ^S473 antibody bead kit (Biosource) according to the protocol presented by the manufacturer.

The experiment shown in FIG. 15 shows that Pim-1 down-regulation decreased baseline phosphorylation of AKT compared to the control, and the effect was significantly enhanced with increasing the concentration of docetaxel. In comparison, BubR1 down-regulation increased baseline phosphorylation of AKT, and the effect was enhanced at lower and not higher concentrations of docetaxel, suggesting that the alternative-signaling pathway may be activated at higher doses. .

The Applicant performed Western blot analysis using antibodies against Pim-1 and BubR1 to measure the protein level at the time point of 48 hours. Equal amounts of protein were loaded on NuPAGE Novex Bis-Tris gels (Invitrogen). Antibodies against Pim-1 (Santa Cruz biotechnology), BubR1 (BD bioscience) and a-tubulin (Sigma) were used as suggested by the manufacturer. The secondary antibody was conjugated to HRP (BioRad), detected by ECL Western Blot Detection System (Amhersham Biosciences), and then exposed to Hyperfilm ECL (Amhersham Biosciences). The experiment shown in FIG. 16 showed that the Pim-1 and BubR1 protein levels were decreased compared to the control, indicating that the decreased protein level corresponded to a functional change (FIG. 15 ). These data suggest that down-regulating Pim-1 protein levels may block phosphorylation and activation of AKT in order to force cells to apoptosis. Pim-1 appears to mediate cell survival by activating pro-survival pathways.

The invention is not limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to be included within the scope of the appended claims.

All documents cited herein are incorporated by reference in their entirety.

<110> AVENTIS PHARMACEUTICALS INC. <120> Method for measuring resistance or sensitivity to docetaxel <130> USAV2004/0088 US PSP <150> US 60/634,298 <151> 2004-12-08 <160> 116 <170> KopatentIn 1.71 <210> 1 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 1 gcgauggaac uucgacuuu 19 <210> 2 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 2 uggaacuucg acuuuguca 19 <210> 3 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 3 agaccaugug gaccuguca 19 <210> 4 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 4 ccuucgaaga aauccagaa 19 <210> 5 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 5 uggugugugg agauauucc 19 <210> 6 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 6 uauuccuuuc gagcaugac 19 <210> 7 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 7 cgaaaggcau guaccauaa 19 <210> 8 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 8 gauacaggcu gaagagauu 19 <210> 9 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 9 gaacaggagc aacuacuaa 19 <210> 10 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 10 gcaaggaccu gaccuacac 19 <210> 11 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 11 gcaaggaccg gaacgagaa 19 <210> 12 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 12 gacccuguca ccaacauuu 19 <210> 13 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 13 caaauccgcu gacugaaau 19 <210> 14 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 14 gaacaagaca gaccgagau 19 <210> 15 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 15 gacccuaugu uuccuaaug 19 <210> 16 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 16 guguauucga caacucugu 19 <210> 17 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 17 gauugcaaga uuggaacaa 19 <210> 18 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 18 guacucaguu gcagacauu 19 <210> 19 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 19 guaucacaaa uuggcuaga 19 <210> 20 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 20 gcacggagau guugcagua 19 <210> 21 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 21 gcaaagaaca ucauccaua 19 <210> 22 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 22 gacaugaaau ccaacaaua 19 <210> 23 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 23 ggaaugagcu ugcaugacu 19 <210> 24 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 24 gagaacugcu acuuauaua 19 <210> 25 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 25 gaauaugcac cacuuggaa 19 <210> 26 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 26 gaaggucgga ugcaugaug 19 <210> 27 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 27 guaaaggaaa guuugguaa 19 <210> 28 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 28 gcaccucgcu ucccacaag 19 <210> 29 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 29 gagcggaccu guaaaacua 19 <210> 30 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 30 gaacgaagag ucacugaag 19 <210> 31 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 31 gcagaaagag gacauauca 19 <210> 32 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 32 gcagcgagcc auugccuuu 19 <210> 33 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 33 gcccgucuau gacaagaaa 19 <210> 34 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 34 ggacguggau ugcaaacaa 19 <210> 35 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 35 gcaugaaugu gcaccauaa 19 <210> 36 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 36 gaaagaacgc uucaacauc 19 <210> 37 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 37 gggcguacca gcagucuga 19 <210> 38 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 38 gcucugggcu caugaacac 19 <210> 39 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 39 gucugaagcc uauaagaug 19 <210> 40 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 40 gagaucgacu cccuguuug 19 <210> 41 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 41 uggaggaguu caagagaaa 19 <210> 42 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 42 gaucaacgac ggagacaag 19 <210> 43 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 43 gaagaaggaa auucaacua 19 <210> 44 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 44 gagaagcguu ucccagugu 19 <210> 45 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 45 gaaacauccu ccggcugaa 19 <210> 46 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 46 gguggaggcu gaugacuug 19 <210> 47 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 47 ggagauugcu gugucuauc 19 <210> 48 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 48 ggagggccau gugaagaug 19 <210> 49 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 49 gaugugaagc ccuccaaug 19 <210> 50 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 50 gcaaugagcc uuuggauau 19 <210> 51 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 51 ggaacaaccu cauucuaaa 19 <210> 52 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 52 ggaagaagau cuagaugua 19 <210> 53 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 53 gaaaucgugg ccgaguucu 19 <210> 54 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 54 guggcauaua uccaucuga 19 <210> 55 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 55 gccgaguucu ucucauucg 19 <210> 56 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 56 ggaacaacug aaagauugg 19 <210> 57 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 57 gucguuacag ucaagcaau 19 <210> 58 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 58 gcacgugacu acuuucaaa 19 <210> 59 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 59 gaugaacuaa gcuugaaua 19 <210> 60 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 60 gagcaguacc acuagaaau 19 <210> 61 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 61 gagcaguacc acuagaaau 19 <210> 62 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 62 gcaccgagac uauuucuug 19 <210> 63 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 63 ggaugaagau gagcuguaa 19 <210> 64 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 64 gcauguggcu cagaucuug 19 <210> 65 <211> 18 <212> RNA <213> Artificial <220> <223> siRNA <400> 65 guacaacagu gaccucca 18 <210> 66 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 66 gaugcuggau gugugaaua 19 <210> 67 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 67 gcuugugaua aagagucaa 19 <210> 68 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 68 gauccaccag augcuauug 19 <210> 69 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 69 gcugucagau aguugauuu 19 <210> 70 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 70 gccuacagcu ucuauaguc 19 <210> 71 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 71 guggacaguu guaaaucua 19 <210> 72 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 72 gaagaucguu uccuauaca 19 <210> 73 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 73 gcacgaaagc aagagauua 19 <210> 74 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 74 gaagaaacca gauggagua 19 <210> 75 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 75 gcuagaaucu gccgucuuu 19 <210> 76 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 76 gaucaaggcu ggagcuuau 19 <210> 77 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 77 gaacaugucg aucaagacu 19 <210> 78 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 78 guuuguaaca gauaucgau 19 <210> 79 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 79 gaguagugca ucgcgaucu 19 <210> 80 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 80 guaacagaua ucgaugaac 19 <210> 81 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 81 gaacgaagaa ugugaucag 19 <210> 82 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 82 guggauuauc acagaucua 19 <210> 83 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 83 gguacaaggc agagcuuaa 19 <210> 84 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 84 gugccacagc agccgacau 19 <210> 85 <211> 21 <212> RNA <213> Artificial <220> <223> siRNA <400> 85 gucgaccauu auguaccgga u 21 <210> 86 <211> 21 <212> RNA <213> Artificial <220> <223> siRNA <400> 86 ggauucguaa cgugauuagc g 21 <210> 87 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 87 gaaacagaag aaccugauu 19 <210> 88 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 88 gaaaggacau gugaacuua 19 <210> 89 <211> 19 <212> RNA <213> Artificial <220> <223> siRNA <400> 89 guacaucuca gaaucuuga 19 <210> 90 <211> 2140 <212> DNA <213> human <400> 90 agctgaggtg tgagcagctg ccgaagtcag ttccttgtgg agccggagct gggcgcggat 60 tcgccgaggc accgaggcac tcagaggagg cgccatgtca gaaccggctg gggatgtccg 120 tcagaaccca tgcggcagca aggcctgccg ccgcctcttc ggcccagtgg acagcgagca 180 gctgagccgc gactgtgatg cgctaatggc gggctgcatc caggaggccc gtgagcgatg 240 gaacttcgac tttgtcaccg agacaccact ggagggtgac ttcgcctggg agcgtgtgcg 300 gggccttggc ctgcccaagc tctaccttcc cacggggccc cggcgaggcc gggatgagtt 360 gggaggaggc aggcggcctg gcacctcacc tgctctgctg caggggacag cagaggaaga 420 ccatgtggac ctgtcactgt cttgtaccct tgtgcctcgc tcaggggagc aggctgaagg 480 gtccccaggt ggacctggag actctcaggg tcgaaaacgg cggcagacca gcatgacaga 540 tttctaccac tccaaacgcc ggctgatctt ctccaagagg aagccctaat ccgcccacag 600 gaagcctgca gtcctggaag cgcgagggcc tcaaaggccc gctctacatc ttctgcctta 660 gtctcagttt gtgtgtctta attattattt gtgttttaat ttaaacacct cctcatgtac 720 ataccctggc cgccccctgc cccccagcct ctggcattag aattatttaa acaaaaacta 780 ggcggttgaa tgagaggttc ctaagagtgc tgggcatttt tattttatga aatactattt 840 aaagcctcct catcccgtgt tctccttttc ctctctcccg gaggttgggt gggccggctt 900 catgccagct acttcctcct ccccacttgt ccgctgggtg gtaccctctg gaggggtgtg 960 gctccttccc atcgctgtca caggcggtta tgaaattcac cccctttcct ggacactcag 1020 acctgaattc tttttcattt gagaagtaaa cagatggcac tttgaagggg cctcaccgag 1080 tgggggcatc atcaaaaact ttggagtccc ctcacctcct ctaaggttgg gcagggtgac 1140 cctgaagtga gcacagccta gggctgagct ggggacctgg taccctcctg gctcttgata 1200 cccccctctg tcttgtgaag gcagggggaa ggtggggtac tggagcagac caccccgcct 1260 gccctcatgg cccctctgac ctgcactggg gagcccgtct cagtgttgag ccttttccct 1320 ctttggctcc cctgtacctt ttgaggagcc ccagcttacc cttcttctcc agctgggctc 1380 tgcaattccc ctctgctgct gtccctcccc cttgtctttc ccttcagtac cctctcatgc 1440 tccaggtggc tctgaggtgc ctgtcccacc cccaccccca gctcaatgga ctggaagggg 1500 aagggacaca caagaagaag ggcaccctag ttctacctca ggcagctcaa gcagcgaccg 1560 ccccctcctc tagctgtggg ggtgagggtc ccatgtggtg gcacaggccc ccttgagtgg 1620 ggttatctct gtgttagggg tatatgatgg gggagtagat ctttctagga gggagacact 1680 ggcccctcaa atcgtccagc gaccttcctc atccacccca tccctcccca gttcattgca 1740 ctttgattag cagcggaaca aggagtcaga cattttaaga tggtggcagt agaggctatg 1800 gacagggcat gccacgtggg ctcatatggg gctgggagta gttgtctttc ctggcactaa 1860 cgttgagccc ctggaggcac tgaagtgctt agtgtacttg gagtattggg gtctgacccc 1920 aaacaccttc cagctcctgt aacatactgg cctggactgt tttctctcgg ctccccatgt 1980 gtcctggttc ccgtttctcc acctagactg taaacctctc gagggcaggg accacaccct 2040 gtactgttct gtgtctttca cagctcctcc cacaatgctg aatatacagc aggtgctcaa 2100 taaatgattc ttagtgactt taaaaaaaaa aaaaaaaaaa 2140 <210> 91 <211> 5449 <212> DNA <213> human <400> 91 gcgccggggg ccgccgcgcc cgccgcccgc tgcctgcgcc gccggccggg catgagttag 60 tcgcagacat ggacaccaaa catttcctgc cgctcgattt ctccacccag gtgaactcct 120 ccctcacctc cccgacgggg cgaggctcca tggctgcccc ctcgctgcac ccgtccctgg 180 ggcctggcat cggctccccg ggacagctgc attctcccat cagcaccctg agctccccca 240 tcaacggcat gggcccgcct ttctcggtca tcagctcccc catgggcccc cactccatgt 300 cggtgcccac cacacccacc ctgggcttca gcactggcag cccccagctc agctcaccta 360 tgaaccccgt cagcagcagc gaggacatca agccccccct gggcctcaat ggcgtcctca 420 aggtccccgc ccacccctca ggaaacatgg cttccttcac caagcacatc tgcgccatct 480 gcggggaccg ctcctcaggc aagcactatg gagtgtacag ctgcgagggg tgcaagggct 540 tcttcaagcg gacggtgcgc aaggacctga cctacacctg ccgcgacaac aaggactgcc 600 tgattgacaa gcggcagcgg aaccggtgcc agtactgccg ctaccagaag tgcctggcca 660 tgggcatgaa gcgggaagcc gtgcaggagg agcggcagcg tggcaaggac cggaacgaga 720 atgaggtgga gtcgaccagc agcgccaacg aggacatgcc ggtggagagg atcctggagg 780 ctgagctggc cgtggagccc aagaccgaga cctacgtgga ggcaaacatg gggctgaacc 840 ccagctcgcc gaacgaccct gtcaccaaca tttgccaagc agccgacaaa cagcttttca 900 ccctggtgga gtgggccaag cggatcccac acttctcaga gctgcccctg gacgaccagg 960 tcatcctgct gcgggcaggc tggaatgagc tgctcatcgc ctccttctcc caccgctcca 1020 tcgccgtgaa ggacgggatc ctcctggcca ccgggctgca cgtccaccgg aacagcgccc 1080 acagcgcagg ggtgggcgcc atctttgaca gggtgctgac ggagcttgtg tccaagatgc 1140 gggacatgca gatggacaag acggagctgg gctgcctgcg cgccatcgtc ctctttaacc 1200 ctgactccaa ggggctctcg aacccggccg aggtggaggc gctgagggag aaggtctatg 1260 cgtccttgga ggcctactgc aagcacaagt acccagagca gccgggaagg ttcgctaagc 1320 tcttgctccg cctgccggct ctgcgctcca tcgggctcaa atgcctggaa catctcttct 1380 tcttcaagct catcggggac acacccattg acaccttcct tatggagatg ctggaggcgc 1440 cgcaccaaat gacttaggcc tgcgggccca tcctttgtgc ccacccgttc tggccaccct 1500 gcctggacgc cagctgttct tctcagcctg agccctgtcc ctgcccttct ctgcctggcc 1560 tgtttggact ttggggcaca gcctgtcact gctctgccta agagatgtgt tgtcaccctc 1620 cttatttctg ttactacttg tctgtggccc agggcagtgg ctttcctgag gcagcagcct 1680 tcgtggcaag aactagcgtg agcccagcca ggcgcctccc caccgggctc tcaggacacc 1740 ctgccacacc ccacggggct tgggcgacta cagggtcttc gggccccagc cctggagctg 1800 caggagttgg gaacggggct tttgtttccg ttgctgttta tcgatgctgg ttttcagaat 1860 tcctgtgtgg ccctcctgtc tggagtgaca tcttcatctg ctctgaatac tggtgcccag 1920 ccagcccgtg acagcttccc cctaatcagg aggggacagc tgggggcgca agctggtgtg 1980 tcatcagcaa agacctcagc cgcctcgggg atgagagggg actcgtgggg caagcaagct 2040 gccctgtgct ctgagtgagg gggaaggtag cccctttttc caaagataac tcacagtttt 2100 gccctcgagc caatgagaac atgagctgcc ctctgtgcaa ggtttcgggg ccacctccag 2160 gctgcagggg cgggtcactc acccccctgt tttctctctg ccttggtgtt ctggtttcag 2220 actcccgact ccccgttcag accagagtgc cccggcccct ccccagcctg agtcttctcc 2280 ttgctctgcg gggtgggctg aggcttgtcc ttgtttcctg cagggctggc cctggctcgg 2340 gcagggtggg gcatcaccac ctcactggcc ttgctggagg cacagggctc tgcggacctg 2400 cagccatctg tgaggcccgc ggggatggga ggggaggagg gtggcctgtt ggtttccctc 2460 agagggggca ggtggcctgg agagagaggg gctcaggaac tgggagcctc gtgggtgggg 2520 cagatgctcc gcggcctgga gtggctctgc cggggcattg gtgggacccc tgctcaggcc 2580 ttctctctgg ctgccagttg tgtctaaaag actcttggaa tctgagaacc cggagtcgca 2640 gcgccctcgg gcctgggcca cacgcaggcc ctggtgggac cacccagcct ggtattgtcc 2700 acggacagcg ttgttcaccc agagccttac ttgggagcct cactgaacgc ctgctctggt 2760 tgaaggtggg gtgggggcgg ggcttggggc ctccctggct cagcccagtg cggcctggcg 2820 ctcctcccgc aggctctgcc cccgggctcc ggtggtgcgg ggccctctca ggttgaactc 2880 gcctcttttg cactggaagg ccctcccttt ggcctgagta cttttcccgt tcacgcctca 2940 gtcccgtgga cccagccttt gtcagtggca ggtgcctgaa cagagggtgg atggggggga 3000 taccggaggg ggtcttgtct tcccagccgc agtctaggaa tgatgcgggg gggtggacgc 3060 cttctccata gtctttcccc acctggagca ggggcttcct cagtggtgag gggagctgcc 3120 tacaggttgg accgggaggc agtggcttgg agaggcagct ttccagcctt ggtggggaag 3180 aaagtgtcca ttctttgcct tcctggagct cccagccaga gctgagctta ggcacccgag 3240 tggagcctgc agctgagtct gtgcccgaga caggctgtca gagattccag aagcctctcc 3300 tccccgccgc cctccacccc tgcctttcag cgttgtggat ccctagaggt ggccccctgc 3360 ccgatccacc gtcctgaggc agagtgttga gcctcatacc tgtaccaggt ccccggccag 3420 ctgggcccct cccaggcact gccaggaagc cccagctgcc cctggcgggt gtggtggaaa 3480 tggcaggagg gtgcaggtac tcttggggcc ccagcggtgg gagtgcaaaa gacccaacgc 3540 caacacctgg tgccttttgc agccagcgcc cacccatccg tgcccggacc cttgggaatg 3600 cccgcggctc cagaggaaaa agcccaggga cggggcctcc gttgcggggg gtcggctgct 3660 tcttgggaac tttgtcgttt ccggcgctgg ctggctggct ggctgtaaag cactgaagcc 3720 ccccggccgc caacccctga aagcagaacc tggcctccct ggccacagca gccttaccca 3780 ccgctctacg tgtcccgggc acttcccgca gccttcccgt ccctttctca tcggccttgt 3840 agttgtacag tgctgttggt ttgaaaaggt gatgtgtggg gagtgcggct catcactgag 3900 tagagaggta gaatttctat ttaaccagac ctgtagtagt attaccaatc cagttcaatt 3960 aaggtgattt tttgtaatta ttattatttt ggtgggacaa tctttaattt tctaaagata 4020 gcactaacat cagctcatta gccacctgtg cctgtccccg ccttggcccg gctggatgaa 4080 gcggcttccc cgcagggccc ccacttccca gtggctgctt cctggggacc cagggcaccc 4140 cggcaccttc aggcacgctc ctcagctggt cacctcccgg ctttgccgtt cagatggggc 4200 tcctgaggct caggagtgaa gatgccacag agccgggctc ccctaggctg cgtcgggcat 4260 gcttggaagc tggcctgcca ggaccttcca ccctggggcc tgtgtcagcc gccggccctc 4320 cgcaccctgg aagcacacgg cctctgggaa ggacagccct gaccttcggt tttccgagca 4380 cggtgtttcc caagaattct gggctggcgg cctggtggca gtgctggaga tgaccccgag 4440 cccctccccg tggggcaccc aggagggccc tgccggaatg tgcagcctgt gggtagtcgg 4500 ctggtgtccc tgtcgtggag ctggggtgcg tgatctggtg ctcgtccacg caggtgtgtg 4560 gtgtaaacat gtatgtgctg tacagagaga cgcgtgtgga gagagccgca caccagcgcc 4620 acccaggaaa ggcggagcgg ttaccagtgt tttgtgttta tttttaatca agacgtttcc 4680 cctgttttcc tataaatttg cttcgtgtaa gcaagtacat aaggaccctc ctttggtgaa 4740 atccgggttc gaatgaatat ctcaaggcag gagatgcatc tattttaaga tgctttggag 4800 cagacagctt tagccgttcc caatccttag caatgcctta gctgggacgc atagctaata 4860 ctttagagag gatgacagat ccataaagag agtaaagata agagaaaatg tctaaagcat 4920 ctggaaaggt aaaaaaaaaa aatctatttt tgtacaaatg taattttatc cctcatgtat 4980 acttggatat ggcgggggga gggctgggac tgtttcgttt ctgcttctag agattgaggt 5040 gaaagcttcg tccgagaaac gccaggacag acgatggcag aggagagggc tcctgtgacg 5100 gcggcgaggc ttgggaggaa accgccgcaa tgggggtgtc ttccctcggg gcaggagggt 5160 gggcctgagg ctttcaaggg ttttcttccc tttcgagtaa tttttaaagc cttgctctgt 5220 tgtgtcctgt tgccggctct ggccttcctg tgactgactg tgaagtggct tctccgtacg 5280 attgtctctg aaacatcgtg gcctcaggtg ccagggtttg atggacagta gcattagaat 5340 tgtggaaaag gaacacgcaa agggagaagt gtgagaggag aaacaaaata tgagcgttta 5400 aaatacatcg ccattcagtt cgttaaaaaa aaaaaaaaaa aaaaaaaaa 5449 <210> 92 <211> 2881 <212> DNA <213> human <400> 92 acagaagtgc tagaagccag tgctcgtgaa ctaaggagaa aaagaacaga caagggaaca 60 gcctggacat ggcatcagag atccacatga caggcccaat gtgcctcatt gagaacacta 120 atgggcgact gatggcgaat ccagaagctc tgaagatcct ttctgccatt acacagccta 180 tggtggtggt ggcaattgtg ggcctctacc gcacaggcaa atcctacctg atgaacaagc 240 tggctggaaa gaaaaagggc ttctctctgg gctccacggt gcagtctcac actaaaggaa 300 tctggatgtg gtgtgtgccc caccccaaga agccaggcca catcctagtt ctgctggaca 360 ccgagggtct gggagatgta gagaagggtg acaaccagaa tgactcctgg atcttcgccc 420 tggccgtcct cctgagcagc accttcgtgt acaatagcat aggaaccatc aaccagcagg 480 ctatggacca actgtactat gtgacagagc tgacacatag aatccgatca aaatcctcac 540 ctgatgagaa tgagaatgag gttgaggatt cagctgactt tgtgagcttc ttcccagact 600 ttgtgtggac actgagagat ttctccctgg acttggaagc agatggacaa cccctcacac 660 cagatgagta cctgacatac tccctgaagc tgaagaaagg taccagtcaa aaagatgaaa 720 cttttaacct gcccagactc tgtatccgga aattcttccc aaagaaaaaa tgctttgtct 780 ttgatcggcc cgttcaccgc aggaagcttg cccagctcga gaaactacaa gatgaagagc 840 tggaccccga atttgtgcaa caagtagcag acttctgttc ctacatcttt agtaattcca 900 aaactaaaac tctttcagga ggcatccagg tcaacgggcc tcgtctagag agcctggtgc 960 tgacctacgt caatgccatc agcagtgggg atctgccgtg catggagaac gcagtcctgg 1020 ccttggccca gatagagaac tcagctgcag tgcaaaaggc tattgcccac tatgaacagc 1080 agatgggcca gaaggtgcag ctgcccacag aaagcctcca ggagctgctg gacctgcaca 1140 gggacagtga gagagaggcc attgaagtct tcatcaggag ttccttcaaa gatgtggacc 1200 atctatttca aaaggagtta gcggcccagc tagaaaaaaa gcgggatgac ttttgtaaac 1260 agaatcagga agcatcatca gatcgttgct caggtttact tcaggtcatt ttcagtcctc 1320 tagaagaaga agtgaaggcg ggaatttatt cgaaaccagg gggctatcgt ctctttgttc 1380 agaagctaca agacctgaag aaaaagtact atgaggaacc gaggaagggg atacaggctg 1440 aagagattct gcagacatac ttgaaatcca aggagtctat gactgatgca attctccaga 1500 cagaccagac tctcacagaa aaagaaaagg agattgaagt ggaacgtgtg aaagctgagt 1560 ctgcacaggc ttcagcaaaa atgttgcagg aaatgcaaag aaagaatgag cagatgatgg 1620 aacagaagga gaggagttat caggaacact tgaaacaact gactgagaag atggagaacg 1680 acagggtcca gttgctgaaa gagcaagaga ggaccctcgc tcttaaactt caggaacagg 1740 agcaactact aaaagaggga tttcaaaaag aaagcagaat aatgaaaaat gagatacagg 1800 atctccagac gaaaatgaga cgacgaaagg catgtaccat aagctaaaga ccagagcctt 1860 cctgtcaccc ctaaccaagg cataattgaa acaattttag aatttggaac aagcgtcact 1920 acatttgata ataattagat cttgcatcat aacaccaaaa gtttataaag gcatgtggta 1980 caatgatcaa aatcatgttt tttcttaaaa aaaaaaaaaa gactgtaaat tgtgcaacaa 2040 agatgcattt acctctgtat caactcagga aatctcataa gctggtacca ctcaggagaa 2100 gtttattctt ccagatgacc agcagtagac aaatggatac tgagcagagt cttaggtaaa 2160 agtcttggga aatatttggg cattggtctg gccaagtcta caatgtccca atatcaagga 2220 caaccaccct agcttcttag tgaagacaat gtacagttat ccattagatc aagactacac 2280 ggtctatgag caataatgtg atttctggac attgcccatg tataatcctc actgatgatt 2340 tcaagctaaa gcaaaccacc ttatacagag atctagaatc tctttatgtt ctccagagga 2400 aggtggaaga aaccatgggc aggagtagga attgagtgat aaacaattgg gctaatgaag 2460 aaaacttctc ttattgttca gttcatccag attataactt caatgggaca ctttagacca 2520 ttagacaatt gacactggat taaacaaatt cacataatgc caaatacaca atgtatttat 2580 agcaacgtat aatttgcaaa gatggacttt aaaagatgct gtgtaactaa actgaaataa 2640 ttcaattact tattatttag aatgttaaag cttatgatag tcttttctaa ttcttaacac 2700 tcatacttga aatctttccg agtttcccca gaagagaata tgggattttt tttgacattt 2760 ttgacccatt taataatgct cttgtgttta cctagtatat gtagactttg tcttatgtgt 2820 caaaagtcct aggaaagtgg ttgatgtttc ttatagcaat taaaaattat ttttgaactg 2880 a 2881 <210> 93 <211> 2684 <212> DNA <213> human <400> 93 cccgagagga gtcggtggca gcggcggcgg cgggaccggc agcagcagca gcagcagcag 60 cagcaaccac tagcctcctg ccccgcggcg ctgccgcacg agccccacga gccgctcacc 120 ccgccgttct cagcgctgcc cgaccccgct ggcgcgccct cccgccgcca gtcccggcag 180 cgccctcagt tgtcctccga ctcgccctcg gccttccgcg ccagccgcag ccacagccgc 240 aacgccaccc gcagccacag ccacagccac agccccaggc atagccttcg gcacagcccc 300 ggctccggct cctgcggcag ctcctctggg caccgtccct gcgccgacat cctggaggtt 360 gggatgctct tgtccaaaat caactcgctt gcccacctgc gcgccgcgcc ctgcaacgac 420 ctgcacgcca ccaagctggc gcccggcaag gagaaggagc ccctggagtc gcagtaccag 480 gtgggcccgc tactgggcag cggcggcttc ggctcggtct actcaggcat ccgcgtctcc 540 gacaacttgc cggtggccat caaacacgtg gagaaggacc ggatttccga ctggggagag 600 ctgcctaatg gcactcgagt gcccatggaa gtggtcctgc tgaagaaggt gagctcgggt 660 ttctccggcg tcattaggct cctggactgg ttcgagaggc ccgacagttt cgtcctgatc 720 ctggagaggc ccgagccggt gcaagatctc ttcgacttca tcacggaaag gggagccctg 780 caagaggagc tggcccgcag cttcttctgg caggtgctgg aggccgtgcg gcactgccac 840 aactgcgggg tgctccaccg cgacatcaag gacgaaaaca tccttatcga cctcaatcgc 900 ggcgagctca agctcatcga cttcgggtcg ggggcgctgc tcaaggacac cgtctacacg 960 gacttcgatg ggacccgagt gtatagccct ccagagtgga tccgctacca tcgctaccat 1020 ggcaggtcgg cggcagtctg gtccctgggg atcctgctgt atgatatggt gtgtggagat 1080 attcctttcg agcatgacga agagatcatc aggggccagg ttttcttcag gcagagggtc 1140 tcttcagaat gtcagcatct cattagatgg tgcttggccc tgagaccatc agataggcca 1200 accttcgaag aaatccagaa ccatccatgg atgcaagatg ttctcctgcc ccaggaaact 1260 gctgagatcc acctccacag cctgtcgccg gggcccagca aatagcagcc tttctggcag 1320 gtcctcccct ctcttgtcag atgcccgagg gaggggaagc ttctgtctcc agcttcccga 1380 gtaccagtga cacgtctcgc caagcaggac agtgcttgat acaggaacaa catttacaac 1440 tcattccaga tcccaggccc ctggaggctg cctcccaaca gtggggaaga gtgactctcc 1500 aggggtccta ggcctcaact cctcccatag atactctctt cttctcatag gtgtccagca 1560 ttgctggact ctgaaatatc ccgggggtgg ggggtggggg tgggtcagaa ccctgccatg 1620 gaactgtttt cttcatcatg agttctgctg aatgccgcga tgggtcaggt aggggggaaa 1680 caggttggga tgggatagga ctagcaccat tttaagtccc tgtcacctct tccgactctt 1740 tctgagtgcc ttctgtgggg actccggctg tgctgggaga aatacttgaa cttgcctctt 1800 ttacctgctg cttctccaaa aatctgcctg ggttttgttc cctatttttc tctcctgtcc 1860 tccctcaccc cctccttcat atgaaaggtg ccatggaaga ggctacaggg ccaaacgctg 1920 agccacctgc ccttttttct gcctccttta gtaaaactcc gagtgaactg gtcttccttt 1980 ttggttttta cttaactgtt tcaaagccaa gacctcacac acacaaaaaa tgcacaaaca 2040 atgcaatcaa cagaaaagct gtaaatgtgt gtacagttgg catggtagta tacaaaaaga 2100 ttgtagtgga tctaattttt aagaaatttt gcctttaagt tattttacct gtttttgttt 2160 cttgttttga aagatgcgca ttctaacctg gaggtcaatg ttatgtattt atttatttat 2220 ttatttggtt cccttcctat tccaagcttc catagctgct gccctagttt tctttcctcc 2280 tttcctcctc tgacttgggg accttttggg ggagggctgc gacgcttgct ctgtttgtgg 2340 ggtgacggga ctcaggcggg acagtgctgc agctccctgg cttctgtggg gcccctcacc 2400 tacttaccca ggtgggtccc ggctctgtgg gtgatgggga ggggcattgc tgactgtgta 2460 tataggataa ttatgaaaag cagttctgga tggtgtgcct tccagatcct ctctggggct 2520 gtgttttgag cagcaggtag cctgctggtt ttatctgagt gaaatactgt acaggggaat 2580 aaaagagatc ttattttttt ttttatactt ggcgtttttt gaataaaaac cttttgtctt 2640 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaa 2684 <210> 94 <211> 2709 <212> DNA <213> human <400> 94 ccctgacccc gcgccgccga gcgccctgag gactcagcga agggtgggcg ccgccgaggc 60 ctcctgccgc tggcgggttt ccgcggagtg ccgcccggct ccgctctgcc gccggcgcgg 120 ctcatgggca gagtcggccg ggcgggccgg cattaaactg aagaaaagat gtccctgtac 180 gatgacctag gagtggagac cagtgactca aaaacagaag gctggtccaa aaacttcaaa 240 cttctgcagt ctcagcttca ggtgaagaag gcagctctca ctcaggcaaa gagccaaagg 300 acgaaacaaa gtacagtcct cgccccagtc attgacctga agcgaggtgg ctcctcagat 360 gaccggcaaa ttgtggacac tccaccgcat gtagcagctg ggctgaagga tcctgttccc 420 agtgggtttt ctgcagggga agttctgatt cccttagctg acgaatatga ccctatgttt 480 cctaatgatt atgagaaagt agtgaagcgc caaagagagg aacgacagag acagcgggag 540 ctggaaagac aaaaggaaat agaagaaagg gaaaaaaggc gtaaagacag acatgaagca 600 agtgggtttg caaggagacc agatccagat tctgatgaag atgaagatta tgagcgagag 660 aggaggaaaa gaagtatggg cggagctgcc attgccccac ccacttctct ggtagagaaa 720 gacaaagagt taccccgaga ttttccttat gaagaggact caagacctcg atcacagtct 780 tccaaagcag ccattcctcc cccagtgtac gaggaacaag acagaccgag atctccaacc 840 ggacctagca actccttcct cgctaacatg gggggcacgg tggcgcacaa gatcatgcag 900 aagtacggct tccgggaggg ccagggtctg gggaagcatg agcagggcct gagcactgcc 960 ttgtcagtgg agaagaccag caagcgtggc ggcaagatca tcgtgggcga cgccacagag 1020 aaagatgcat ccaagaagtc agattcaaat ccgctgactg aaatacttaa gtgtcctact 1080 aaagtggtct tactaaggaa catggttggt gcgggagagg tggatgaaga cttggaagtt 1140 gaaaccaagg aagaatgtga aaaatatggc aaagttggaa aatgtgtgat atttgaaatt 1200 cctggtgccc ctgatgatga agcagtacgg atatttttag aatttgagag agttgaatca 1260 gcaattaaag cggttgttga cttgaatggg aggtattttg gtggacgggt ggtaaaagca 1320 tgtttctaca atttggacaa attcagggtc ttggatttgg cagaacaagt ttgattttaa 1380 gaactagagc acgagtcatc tccggtgatc cttaaatgaa ctgcaggctg agaaaagaag 1440 gaaaaaggtc acagcctcca tggctgttgc ataccaagac tcttggaagg acttctaaga 1500 tatatgttga ttgatccctt ttttattttg tggtttttta atatagtata aaaatccttt 1560 taaaaaaaca acaatctgtg tgcctctctg gttgtttctc ttttttatta ttactcctga 1620 gttgatgaca ttttttgtta gatttcatgg taattctcaa gtgcttcaat gatgcagcat 1680 ttcttgcact aaaaaaaaaa aaaaaaaaaa aactagaaag ttttgggaca tggggttata 1740 ttaaattatt ctttgttttt ctttttcttt taataaagcc tgcaagttac taaattgtag 1800 tttcataaat tctgtagtaa agtatcatct tggcagtgtg ccaaaggtga aaatgatgct 1860 ttctctaaca gagaaattct tagtgactcc agtcgtagaa aaacgtcttt acaacctgaa 1920 taagattgaa gaattgtgaa cataccatgg cctattggat gaatcatttg ccgtaggcta 1980 aatcagactg tagggtttgt gatggattta tggagtatgt gggtatagaa atcatgaatc 2040 tagcatttgt tttcagagat tcaagcatag tcttaagggt agatcagaaa tgacaaatga 2100 attcaaaacc tagcaggtgc attgtaaatg tgtgcccagt tatgttttgg aaatggcagt 2160 tccttggggt catgtttcta ctggcaaaat ttgcaatagt gttctattgt atgtaatttt 2220 aaaatttata agattatcca cgttggccaa gtaaactgta ctgccaatag aattctggaa 2280 ttgtgagaaa ttgtatcatt gaagttcagt aggatgtgtg gcttaaaaat ttatcaggac 2340 cacaaaaaag aaaacaaaaa tatttggtac tgaggttcat tgccagggca ggaggtattt 2400 ccagaaaata ctcatgcctg tgttctgttc cttgctttcc caaatactgc atgtgacttt 2460 cctaagcggc agctgaaaga ctcgagcccg tgctgtctcc tttggttatt atgacatgaa 2520 agtgtatcaa gaactcagca tttctttgca tccatggact tggtttggag acataaggaa 2580 tattctgacc ctttttaaaa aaggattttc tcatgttttt atttaacata aataaaagaa 2640 taacatttta tcttttgtgg tattatttta ttgaataaaa ttgagtttta tgataaaaaa 2700 aaaaaaaaa 2709 <210> 95 <211> 2150 <212> DNA <213> human <400> 95 ctcgagccac gaaggccccg ctgtcctgtc tagcagatac ttgcacggtt tacagaaatt 60 cggtccctgg gtcgtgtcag gaaactggaa aaaaggtcat aagcatgaag cgcagttcag 120 tttccagcgg tggtgctggc cgcctctcca tgcaggagtt aagatcccag gatgtaaata 180 aacaaggcct ctatacccct caaaccaaag agaaaccaac ctttggaaag ttgagtataa 240 acaaaccgac atctgaaaga aaagtctcgc tatttggcaa aagaactagt ggacatggat 300 cccggaatag tcaacttggt atattttcca gttctgagaa aatcaaggac ccgagaccac 360 ttaatgacaa agcattcatt cagcagtgta ttcgacaact ctgtgagttt cttacagaaa 420 atggttatgc acataatgtg tccatgaaat ctctacaagc tccctctgtt aaagacttcc 480 tgaagatctt cacatttctt tatggcttcc tgtgcccctc atacgaactt cctgacacaa 540 agtttgaaga agaggttcca agaatcttta aagaccttgg gtatcctttt gcactatcca 600 aaagctccat gtacacagtg ggggctcctc atacatggcc tcacattgtg gcagccttag 660 tttggctaat agactgcatc aagatacata ctgccatgaa agaaagctca cctttatttg 720 atgatgggca gccttgggga gaagaaactg aagatggaat tatgcataat aagttgtttt 780 tggactacac cataaaatgc tatgagagtt ttatgagtgg tgccgacagc tttgatgaga 840 tgaatgcaga gctgcagtca aaactgaagg atttatttaa tgtggatgct tttaagctgg 900 aatcattaga agcaaaaaac agagcattga atgaacagat tgcaagattg gaacaagaaa 960 gagaaaaaga accgaatcgt ctagagtcgt tgagaaaact gaaggcttcc ttacaaggag 1020 atgttcaaaa gtatcaggca tacatgagca atttggagtc tcattcagcc attcttgacc 1080 agaaattaaa tggtctcaat gaggaaattg ctagagtaga actagaatgt gaaacaataa 1140 aacaggagaa cactcgacta cagaatatca ttgacaacca gaagtactca gttgcagaca 1200 ttgagcgaat aaatcatgaa agaaatgaat tgcagcagac tattaataaa ttaaccaagg 1260 acctggaagc tgaacaacag aagttgtgga atgaggagtt aaaatatgcc agaggcaaag 1320 aagcgattga aacacaatta gcagagtatc acaaattggc tagaaaatta aaacttattc 1380 ctaaaggtgc tgagaattcc aaaggttatg actttgaaat taagtttaat cccgaggctg 1440 gtgccaactg ccttgtcaaa tacagggctc aagtttatgt acctcttaag gaactcctga 1500 atgaaactga agaagaaatt aataaagccc taaataaaaa aatgggtttg gaggatactt 1560 tagaacaatt gaatgcaatg ataacagaaa gcaagagaag tgtgagaact ctgaaagaag 1620 aagttcaaaa gctggatgat ctttaccaac aaaaaattaa ggaagcagag gaagaggatg 1680 aaaaatgtgc cagtgagctt gagtccttgg agaaacacaa gcacctgcta gaaagtactg 1740 ttaaccaggg gctcagtgaa gctatgaatg aattagatgc tgttcagcgg gaataccaac 1800 tagttgtgca aaccacgact gaagaaagac gaaaagtggg aaataacttg caacgtctgt 1860 tagagatggt tgctacacat gttgggtctg tagagaaaca tcttgaggag cagattgcta 1920 aagttgatag agaatatgaa gaatgcatgt cagaagatct ctcggaaaat attaaagaga 1980 ttagagataa gtatgagaag aaagctactc taattaagtc ttctgaagaa tgaagataaa 2040 atgttgatca tgtatatata tccatagtga ataaaattgt ctcagtaaaa aaaaaaaaaa 2100 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 2150 <210> 96 <211> 2977 <212> DNA <213> human <400> 96 ccgaatgtga ccgcctcccg ctccctcacc cgccgcgggg aggaggagcg ggcgagaagc 60 tgccgccgaa cgacaggacg ttggggcggc ctggctccct caggtttaag aattgtttaa 120 gctgcatcaa tggagcacat acagggagct tggaagacga tcagcaatgg ttttggattc 180 aaagatgccg tgtttgatgg ctccagctgc atctctccta caatagttca gcagtttggc 240 tatcagcgcc gggcatcaga tgatggcaaa ctcacagatc cttctaagac aagcaacact 300 atccgtgttt tcttgccgaa caagcaaaga acagtggtca atgtgcgaaa tggaatgagc 360 ttgcatgact gccttatgaa agcactcaag gtgaggggcc tgcaaccaga gtgctgtgca 420 gtgttcagac ttctccacga acacaaaggt aaaaaagcac gcttagattg gaatactgat 480 gctgcgtctt tgattggaga agaacttcaa gtagatttcc tggatcatgt tcccctcaca 540 acacacaact ttgctcggaa gacgttcctg aagcttgcct tctgtgacat ctgtcagaaa 600 ttcctgctca atggatttcg atgtcagact tgtggctaca aatttcatga gcactgtagc 660 accaaagtac ctactatgtg tgtggactgg agtaacatca gacaactctt attgtttcca 720 aattccacta ttggtgatag tggagtccca gcactacctt ctttgactat gcgtcgtatg 780 cgagagtctg tttccaggat gcctgttagt tctcagcaca gatattctac acctcacgcc 840 ttcaccttta acacctccag tccctcatct gaaggttccc tctcccagag gcagaggtcg 900 acatccacac ctaatgtcca catggtcagc accacgctgc ctgtggacag caggatgatt 960 gaggatgcaa ttcgaagtca cagcgaatca gcctcacctt cagccctgtc cagtagcccc 1020 aacaatctga gcccaacagg ctggtcacag ccgaaaaccc ccgtgccagc acaaagagag 1080 cgggcaccag tatctgggac ccaggagaaa aacaaaatta ggcctcgtgg acagagagat 1140 tcaagctatt attgggaaat agaagccagt gaagtgatgc tgtccactcg gattgggtca 1200 ggctcttttg gaactgttta taagggtaaa tggcacggag atgttgcagt aaagatccta 1260 aaggttgtcg acccaacccc agagcaattc caggccttca ggaatgaggt ggctgttctg 1320 cgcaaaacac ggcatgtgaa cattctgctt ttcatggggt acatgacaaa ggacaacctg 1380 gcaattgtga cccagtggtg cgagggcagc agcctctaca aacacctgca tgtccaggag 1440 accaagtttc agatgttcca gctaattgac attgcccggc agacggctca gggaatggac 1500 tatttgcatg caaagaacat catccataga gacatgaaat ccaacaatat atttctccat 1560 gaaggcttaa cagtgaaaat tggagatttt ggtttggcaa cagtaaagtc acgctggagt 1620 ggttctcagc aggttgaaca acctactggc tctgtcctct ggatggcccc agaggtgatc 1680 cgaatgcagg ataacaaccc attcagtttc cagtcggatg tctactccta tggcatcgta 1740 ttgtatgaac tgatgacggg ggagcttcct tattctcaca tcaacaaccg agatcagatc 1800 atcttcatgg tgggccgagg atatgcctcc ccagatctta gtaagctata taagaactgc 1860 cccaaagcaa tgaagaggct ggtagctgac tgtgtgaaga aagtaaagga agagaggcct 1920 ctttttcccc agatcctgtc ttccattgag ctgctccaac actctctacc gaagatcaac 1980 cggagcgctt ccgagccatc cttgcatcgg gcagcccaca ctgaggatat caatgcttgc 2040 acgctgacca cgtccccgag gctgcctgtc ttctagttga ctttgcacct gtcttcaggc 2100 tgccagggga ggaggagaag ccagcaggca ccacttttct gctccctttc tccagaggca 2160 gaacacatgt tttcagagaa gctctgctaa ggaccttcta gactgctcac agggccttaa 2220 cttcatgttg ccttcttttc tatccctttg ggccctggga gaaggaagcc atttgcagtg 2280 ctggtgtgtc ctgctccctc cccacattcc ccatgctcaa ggcccagcct tctgtagatg 2340 cgcaagtgga tgttgatggt agtacaaaaa gcaggggccc agccccagct gttggctaca 2400 tgagtattta gaggaagtaa ggtagcaggc agtccagccc tgatgtggag acacatggga 2460 ttttggaaat cagcttctgg aggaatgcat gtcacaggcg ggactttctt cagagagtgg 2520 tgcagcgcca gacattttgc acataaggca ccaaacagcc caggactgcc gagactctgg 2580 ccgcccgaag gagcctgctt tggtactatg gaacttttct taggggacac gtcctccttt 2640 cacagcttct aaggtgtcca gtgcattggg atggttttcc aggcaaggca ctcggccaat 2700 ccgcatctca gccctctcag gagcagtctt ccatcatgct gaattttgtc ttccaggagc 2760 tgcccctatg gggcgggccg cagggccagc ctgtttctct aacaaacaaa caaacaaaca 2820 gccttgtttc tctagtcaca tcatgtgtat acaaggaagc caggaataca ggttttcttg 2880 atgatttggg ttttaatttt gtttttattg cacctgacaa aatacagtta tctgatggtc 2940 cctcaattat gttattttaa taaaataaat taaattt 2977 <210> 97 <211> 1212 <212> DNA <213> human <400> 97 atggaccgat ctaaagaaaa ctgcatttca ggacctgtta aggctacagc tccagttgga 60 ggtccaaaac gtgttctcgt gactcagcaa tttccttgtc agaatccatt acctgtaaat 120 agtggccagg ctcagcgggt cttgtgtcct tcaaattctt cccagcgcat tcctttgcaa 180 gcacaaaagc ttgtctccag tcacaagccg gttcagaatc agaagcagaa gcaattgcag 240 gcaaccagtg tacctcatcc tgtctccagg ccactgaata acacccaaaa gagcaagcag 300 cccctgccat cggcacctga aaataatcct gaggaggaac tggcatcaaa acagaaaaat 360 gaagaatcaa aaaagaggca gtgggctttg gaagactttg aaattggtcg ccctctgggt 420 aaaggaaagt ttggtaatgt ttatttggca agagaaaagc aaagcaagtt tattctggct 480 cttaaagtgt tatttaaagc tcagctggag aaagccggag tggagcatca gctcagaaga 540 gaagtagaaa tacagtccca ccttcggcat cctaatattc ttagactgta tggttatttc 600 catgatgcta ccagagtcta cctaattctg gaatatgcac cacttggaac agtttataga 660 gaacttcaga aactttcaaa gtttgatgag cagagaactg ctacttatat aacagaattg 720 gcaaatgccc tgtcttactg ccattcgaag agagttattc atagagacat taagccagag 780 aacttacttc ttggatcagc tggagagctt aaaattgcag attttgggtg gtcagtacat 840 gctccatctt ccaggaggac cactctctgt ggcaccctgg actacctgcc ccctgaaatg 900 attgaaggtc ggatgcatga tgagaaggtg gatctctgga gccttggagt tctttgctat 960 gaatttttag ttgggaagcc tccttttgag gcaaacacat accaagagac ctacaaaaga 1020 atatcacggg ttgaattcac attccctgac tttgtaacag agggagccag ggacctcatt 1080 tcaagactgt tgaagcataa tcccagccag aggccaatgc tcagagaagt acttgaacac 1140 ccctggatca cagcaaattc atcaaaacca tcaaattgcc aaaacaaaga atcagctagc 1200 aaacagtctt ag 1212 <210> 98 <211> 2788 <212> DNA <213> human <400> 98 ggcggcggta gcagccaggc ttggcccccg gcgtggagca gacgcggacc cctccttcct 60 ggcggcggcg gcgcgggctc agagcccggc aacgggcggg cgggcagaat gagtctgcag 120 gtcttaaacg acaaaaatgt cagcaatgaa aaaaatacag aaaattgcga cttcctgttt 180 tcgccaccag aagttaccgg aagatcgtct gttcttcgtg tgtcacagaa agaaaatgtg 240 ccacccaaga acctggccaa agctatgaag gtgacttttc agacacctct gcgggatcca 300 cagacgcaca ggattctaag tcctagcatg gccagcaaac ttgaggctcc tttcactcag 360 gatgacaccc ttggactgga aaactcacac ccggtctgga cacagaaaga gaaccaacag 420 ctcatcaagg aagtggatgc caaaactact catggaattc tacagaaacc agtggaggct 480 gacaccgacc tcctggggga tgcaagccca gcctttggga gtggcagctc cagcgagtct 540 ggcccaggtg ccctggctga cctggactgc tcaagctctt cccagagccc aggaagttct 600 gagaaccaaa tggtgtctcc aggaaaagtg tctggcagcc ctgagcaagc cgtggaggaa 660 aaccttagtt cctattcctt agacagaaga gtgacacccg cctctgagac cctagaagac 720 ccttgcagga cagagtccca gcacaaagcg gagactccgc acggagccga ggaagaatgc 780 aaagcggaga ctccgcacgg agccgaggag gaatgccggc acggtggggt ctgtgctccc 840 gcagcagtgg ccacttcgcc tcctggtgca atccctaagg aagcctgcgg aggagcaccc 900 ctgcagggtc tgcctggcga agccctgggc tgccctgcgg gtgtgggcac ccccgtgcca 960 gcagatggca ctcagaccct tacctgtgca cacacctctg ctcctgagag cacagcccca 1020 accaaccacc tggtggctgg cagggccatg accctgagtc ctcaggaaga agtggctgca 1080 ggccaaatgg ccagctcctc gaggagcgga cctgtaaaac tagaatttga tgtatctgat 1140 ggcgccacca gcaaaagggc acccccacca aggagactgg gagagaggtc cggcctcaag 1200 cctcccttga ggaaagcagc agtgaggcag caaaaggccc cgcaggaggt ggaggaggac 1260 gacggtagga gcggagcagg agaggacccc cccatgccag cttctcgggg ctcttaccac 1320 ctcgactggg acaaaatgga tgacccaaac ttcatcccgt tcggaggtga caccaagtct 1380 ggttgcagtg aggcccagcc cccagaaagc cctgagacca ggctgggcca gccagcggct 1440 gaacagttgc atgctgggcc tgccacggag gagccaggtc cctgtctgag ccagcagctg 1500 cattcagcct cagcggagga cacgcctgtg gtgcagttgg cagccgagac cccaacagca 1560 gagagcaagg agagagcctt gaactctgcc agcacctcgc ttcccacaag ctgtccaggc 1620 agtgagccag tgcccaccca tcagcagggg cagcctgcct tggagctgaa agaggagagc 1680 ttcagagacc ccgctgaggt tctaggcacg ggcgcggagg tggattacct ggagcagttt 1740 ggaacttcct cgtttaagga gtcggccttg aggaagcagt ccttatacct caagttcgac 1800 cccctcctga gggacagtcc tggtagacca gtgcccgtgg ccaccgagac cagcagcatg 1860 cacggtgcaa atgagactcc ctcaggacgt ccgcgggaag ccaagcttgt ggagttcgat 1920 ttcttgggag cactggacat tcctgtgcca ggcccacccc caggtgttcc cgcgcctggg 1980 ggcccacccc tgtccaccgg acctatagtg gacctgctcc agtacagcca gaaggacctg 2040 gatgcagtgg taaaggcgac acaggaggag aaccgggagc tgaggagcag gtgtgaggag 2100 ctccacggga agaacctgga actggggaag atcatggaca ggttcgaaga ggttgtgtac 2160 caggccatgg aggaagttca gaagcagaag gaactttcca aagctgaaat ccagaaagtt 2220 ctaaaagaaa aagaccaact taccacagat ctgaactcca tggagaagtc cttctccgac 2280 ctcttcaagc gttttgagaa acagaaagag gtgatcgagg gctaccgcaa gaacgaagag 2340 tcactgaaga agtgcgtgga ggattacctg gcaaggatca cccaggaggg ccagaggtac 2400 caagccctga aggcccacgc ggaggagaag ctgcagctgg caaacgagga gatcgcccag 2460 gtccggagca aggcccaggc ggaagcgttg gccctccagg ccagcctgag gaaggagcag 2520 atgcgcatcc agtcgctgga gaagacagtg gagcagaaga ctaaagagaa cgaggagctg 2580 accaggatct gcgacgacct catctccaag atggagaaga tctgacctcc acggagccgc 2640 tgtccccgcc cccctgctcc cgtctgtctg tcctgtctga ttctcttagg tgtcatgttc 2700 ttttttctgt cttgtcttca acttttttta aaactagatt gctttgaaaa catgactcaa 2760 taaaagtttc ctttcaattt aaaaaaaa 2788 <210> 99 <211> 2287 <212> DNA <213> human <400> 99 cgcgccccgc gcagccccgg gcgccgcgcg tcctgcccgg cctgcggccc cagcccttgc 60 gccgctcgtc cgacccgcga tcgtccacca gaccgtgcct cccggccgcc cggccggccc 120 gcgtgcatgc ttcggtctgg gccagcctct gggccgtccg tccccactgg ccgggccatg 180 ccgagtcgcc gcgtcgccag accgccggct gcgccggagc tgggggcctt agggtccccc 240 gacctctcct cactctcgct cgccgtttcc aggagcacag atgaattgga gatcatcgac 300 gagtacatca aggagaacgg cttcggcctg gacgggggac agccgggccc gggcgagggg 360 ctgccacgcc tggtgtctcg cggggctgcg tccctgagca cggtcaccct gggccctgtg 420 gcgcccccag ccacgccgcc gccttggggc tgccccctgg gccgactagt gtccccagcg 480 ccgggcccgg gcccgcagcc gcacctggtc atcacggagc agcccaagca gcgcggcatg 540 cgcttccgct acgagtgcga gggccgctcg gccggcagca tccttgggga gagcagcacc 600 gaggccagca agacgctgcc cgccatcgag ctccgggatt gtggagggct gcgggaggtg 660 gaggtgactg cctgcctggt gtggaaggac tggcctcacc gagtccaccc ccacagcctc 720 gtggggaaag actgcaccga cggcatctgc agggtgcggc tccggcctca cgtcagcccc 780 cggcacagtt ttaacaacct gggcatccag tgtgtgagga agaaggagat tgaggctgcc 840 attgagcgga agattcaact gggcattgac ccctacaacg ctgggtccct gaagaaccat 900 caggaagtag acatgaatgt ggtgaggatc tgcttccagg cctcatatcg ggaccagcag 960 ggacagatgc gccggatgga tcctgtgctt tccgagcccg tctatgacaa gaaatccaca 1020 aacacatcag agctgcggat ttgccgaatt aacaaggaaa gcgggccgtg caccggtggc 1080 gaggagctct acttgctctg cgacaaggtg cagaaagagg acatatcagt ggtgttcagc 1140 agggcctcct gggaaggtcg ggctgacttc tcccaggccg acgtgcaccg ccagattgcc 1200 attgtgttca agacgccgcc ctacgaggac ctggagattg tcgagcccgt gacagtcaac 1260 gtcttcctgc agcggctcac cgatggggtc tgcagcgagc cattgccttt cacgtacctg 1320 cctcgcgacc atgacagcta cggcgtggac aagaagcgga aacgggggat gcccgacgtc 1380 cttggggagc tgaacagctc tgacccccat ggcatcgaga gcaaacggcg gaagaaaaag 1440 ccggccatcc tggaccactt cctgcccaac cacggctcag gcccgttcct cccgccgtca 1500 gccctgctgc cagaccctga cttcttctct ggcaccgtgt ccctgcccgg cctggagccc 1560 cctggcgggc ctgacctcct ggacgatggc tttgcctacg accctacggc ccccacactc 1620 ttcaccatgc tggacctgct gcccccggca ccgccacacg ctagcgctgt tgtgtgcagc 1680 ggaggtgccg gggccgtggt tggggagacc cccggccctg aaccactgac actggactcg 1740 taccaggccc cgggccccgg ggatggaggc accgccagcc ttgtgggcag caacatgttc 1800 cccaatcatt accgcgaggc ggcctttggg ggcggcctcc tatccccggg gcctgaagcc 1860 acgtagcccc gcgatgccag aggaggggca ctgggtgggg agggaggtgg aggagccgtg 1920 caatcccaac caggatgtct agcaccccca tccccttggc ccttcctcat gcttctgaag 1980 tggacatatt cagccttggc gagaagctcc gttgcacggg tttccccttg agcccatttt 2040 acagatgagg aaactgagtc cggagaggaa aagggacatg gctcccgtgc actagcttgt 2100 tacagctgcc tctgtcccca catgtggggg caccttctcc agtaggattc ggaaaagatt 2160 gtacatatgg gaggaggggg cagattcctg gccctccctc cccagacttg aaggtggggg 2220 gtaggttggt tgttcagagt cttcccaata aagatgagtt tttgagcctc aaaaaaaaaa 2280 aaaaaaa 2287 <210> 100 <211> 2850 <212> DNA <213> human <400> 100 tcgcacttcc gtgtgccgcg gcgccggagc ccgaggcggc tgtagcccac atctcccgag 60 cgacccccgg cgcccgcccg ccgcgcggag gcccgggcca cacctcactg gccgcttggc 120 ccatcccagt cagcgccgcg ccgaaccccg tccgcgcgcg ccggggagcg gcgcccccgc 180 cgctgccgcc gcgacccttg gcgcctgccc ctgcaacggg aggtctgcag ggaactggcc 240 aggcaagggg gcaggcccgt ttctcctggt ggttggtgcg ttgtagcagc agcgggagcc 300 aggactaagg acaagcagga gctgggagcc ccagccccac tgcaggcccc accatggcgc 360 cgttcctgcg catcgccttc aactcctatg agctgggctc cctgcaggcc gaggacgagg 420 cgaaccagcc cttctgtgcc gtgaagatga aggaggcgct cagcacagag cgtgggaaaa 480 cactggtgca gaagaagccg accatgtatc ctgagtggaa gtcgacgttc gatgcccaca 540 tctatgaggg gcgcgtcatc cagattgtgc taatgcgggc agcagaggag ccagtgtctg 600 aggtgaccgt gggtgtgtcg gtgctggccg agcgctgcaa gaagaacaat ggcaaggctg 660 agttctggct ggacctgcag cctcaggcca aggtgttgat gtctgttcag tatttcctgg 720 aggacgtgga ttgcaaacag tctatgcgca gtgaggacga ggccaagttc ccaacgatga 780 accgccgcgg agccatcaaa caggccaaaa tccactacat caagaaccat gagtttatcg 840 ccaccttctt tgggcaaccc accttctgtt ctgtgtgcaa agactttgtc tggggcctca 900 acaagcaagg ctacaaatgc aggcaatgta acgctgccat ccacaagaaa tgcatcgaca 960 agatcatcgg cagatgcact ggcaccgcgg ccaacagccg ggacactata ttccagaaag 1020 aacgcttcaa catcgacatg ccgcaccgct tcaaggttca caactacatg agccccacct 1080 tctgtgacca ctgcggcagc ctgctctggg gactggtgaa gcagggatta aagtgtgaag 1140 actgcggcat gaatgtgcac cataaatgcc gggagaaggt ggccaacctc tgcggcatca 1200 accagaagct tttggctgag gccttgaacc aagtcaccca gagagcctcc cggagatcag 1260 actcagcctc ctcagagcct gttgggatat atcagggttt cgagaagaag accggagttg 1320 ctggggagga catgcaagac aacagtggga cctacggcaa gatctgggag ggcagcagca 1380 agtgcaacat caacaacttc atcttccaca aggtcctggg caaaggcagc ttcgggaagg 1440 tgctgcttgg agagctgaag ggcagaggag agtactttgc catcaaggcc ctcaagaagg 1500 atgtggtcct gatcgacgac gacgtggagt gcaccatggt tgagaagcgg gtgctgacac 1560 ttgccgcaga gaatcccttt ctcacccacc tcatctgcac cttccagacc aaggaccacc 1620 tgttctttgt gatggagttc ctcaacgggg gggacctgat gtaccacatc caggacaaag 1680 gccgctttga actctaccgt gccacgtttt atgccgctga gataatgtgt ggactgcagt 1740 ttctacacag caagggcatc atttacaggg acctcaaact ggacaatgtg ctgctggacc 1800 gggatggcca catcaagatt gccgactttg ggatgtgcaa agagaacata ttcggggaga 1860 gccgggccag caccttctgc ggcacccctg actatatcgc ccctgagatc ctacagggcc 1920 tgaagtacac attctctgtg gactggtggt ctttcggggt ccttctgtac gagatgctca 1980 ttggccagtc ccccttccat ggtgatgatg aggatgaact cttcgagtcc atccgtgtgg 2040 acacgccaca ttatccccgc tggatcacca aggagtccaa ggacatcctg gagaagctct 2100 ttgaaaggga accaaccaag aggctgggag tgaccggaaa catcaaaatc caccccttct 2160 tcaagaccat aaactggact ctgctggaaa agcggaggtt ggagccacct ttcaggccca 2220 aagtgaagtc acccagagac tacagtaact ttgaccagga gttcctgaac gagaaggcgc 2280 gcctctccta cagcgacaag aacctcatcg actccatgga ccagtctgca ttcgctggct 2340 tctcctttgt gaaccccaaa ttcgagcacc tcctggaaga ttgaggttcc tggacagatc 2400 aggctagccc tgccctccac ccacacctgc ccgctcccca cgataagcac cagtgggact 2460 gtggtgactt ctgctgctgg ccccgcccct gcccccagag cgtccttggc tgccgtctgg 2520 ccgggctctc atggtacttc ctctgtgaac tgtgtgtgaa tctgcttttc ctctgccttc 2580 ggagggaaat tgtaaatcct gtgtttcatt acttgaatgt agttatctat tgaaaatata 2640 tattatatac atagacatat atatatatat aataggctgt atatattgct cagtagagaa 2700 aaaccatggg ggactggtga tatgttgatc tttttcaaaa aaatatatat atgacaaaaa 2760 aaaaaaaaaa ggagcacaag ctgtttgaac caccaggttt atttgtgtgt ctaaataaac 2820 accaaatagt accaaaaaaa aaaaaaaaaa 2850 <210> 101 <211> 1584 <212> DNA <213> human <400> 101 gttggttgga gcgtcgcgca gtcgggaggt ccgggaaagt ttctttggag gtccggcccg 60 gagcggccat gtcccacggc cccaagcagc ccggcgcggc cgccgcgccg gcgggcggca 120 aggctccggg ccagcatggg ggcttcgtgg tgactgtcaa gcaagagcgc ggcgagggtc 180 cacgcgcggg cgagaagggg tcccacgagg aggagccggt gaagaaacgc ggctggccca 240 agggcaagaa gcggaagaag attctgccga atgggcccaa ggcaccggtc acgggctacg 300 tgcgcttcct gaacgagcgg cgcgagcaga tccgcacgcg ccacccggat ctgccctttc 360 ccgagatcac caagatgctg ggcgccgagt ggagcaagct gcagccaacg gaaaagcagc 420 ggtacctgga tgaggccgag agagagaagc agcagtacat gaaggagctg cgggcgtacc 480 agcagtctga agcctataag atgtgcacgg agaagatcca ggagaagaag atcaagaaag 540 aagactcgag ctctgggctc atgaacactc tcctgaatgg acacaagggt ggggactgcg 600 atggcttctc caccttcgat gttcccatct tcactgaaga gttcttggac caaaacaaag 660 cgcgtgaggc ggagcttcgg cgcttgcgga agatgaatgt ggccttcgag gagcagaacg 720 cggtactgca gaggcacacg cagagcatga gcagcgcgcg cgagcgtctg gagcaggagc 780 tggcgctgga ggagcggagg acgctggcgc tgcagcagca gctccaggcc gtgcgccagg 840 cgctcaccgc cagcttcgcc tcactgccgg tgccgggcac gggcgaaacg cccacgctgg 900 gcactctgga cttctacatg gcccggcttc acggagccat cgagcgcgac cccgcccagc 960 acgagaagct catcgtccgc atcaaggaaa tcctggccca ggtcgccagc gagcacctgt 1020 gaggagtggg cgggcccacg atgcagagga gaagctgtgg gcgcggccct gccacacccc 1080 accccgtgga cgagaggctg ggggtccacc ctttggggcc tggtcccatc ctgcaccttg 1140 ggggctccag cccccctaaa attaaatttc tgcagcatcc ctttagcttt caatctcccc 1200 agccccctga acccggaaaa agcactcgct gcgcgataca cccagaagaa cctcacagcc 1260 gagggtgccc ctcctcggag gacagccacg cgctacactg gctctccggg ccacccccag 1320 gacacagggc agacgaaacc cacccccagc acacggcagg accccccaaa ttactcacta 1380 cggggggctg tgccataggc cacacaggaa gctgccttgt ggggacttac ctggggtgtc 1440 ccccgcatgc ctgtacccca gatgggtggg ggccggcttt gcccatcctg ctctcctcca 1500 gccgagggac cctggtgggg gtggctcctt ctcactgctg gatccggact ttttaaataa 1560 aaacaagtaa aatttgtgtt ttaa 1584 <210> 102 <211> 2383 <212> DNA <213> human <400> 102 gctgctgcga cagtccacta cctttttcga gagtgactcc cgttgtccca aggcttccca 60 gagcgaacct gtgcggctgc aggcaccggc gcgtcgagtt tccggcgtcc ggaaggaccg 120 agctcttctc gcggatccag tgttccgttt ccagccccca atctcagagc cgagccgaca 180 gagagcaggg aaccggcatg gccaaagccg cggcgatcgg catcgacctg ggcaccacct 240 actcctgcgt gggggtgttc caacacggca aggtggagat catcgccaac gaccagggca 300 accgcaccac ccccagctac gtggccttca cggacaccga gcggctcatc ggggatgcgg 360 ccaagaacca ggtggcgctg aacccgcaga acaccgtgtt tgacgcgaag cggctgatcg 420 gccgcaagtt cggcgacccg gtggtgcagt cggacatgaa gcactggcct ttccaggtga 480 tcaacgacgg agacaagccc aaggtgcagg tgagctacaa gggggacacc aaggcattct 540 accccgagga gatctcgtcc atggtgctga ccaagatgaa ggagatcgcc gaggcgtacc 600 tgggctaccc ggtgaccaac gcggtgatca ccgtgccggc ctacttcaac gactcgcagc 660 gccaggccac caaggatgcg ggtgtgatcg cggggctcaa cgtgctgcgg atcatcaacg 720 agcccacggc cgccgccatc gcctacggcc tggacagaac gggcaagggg gagcgcaacg 780 tgctcatctt tgacctgggc gggggcacct tcgacgtgtc catcctgacg atcgacgacg 840 gcatcttcga ggtgaaggcc acggccgggg acacccacct gggtggggag gactttgaca 900 acaggctggt gaaccacttc gtggaggagt tcaagagaaa acacaagaag gacatcagcc 960 agaacaagcg agccgtgagg cggctgcgca ccgcctgcga gagggccaag aggaccctgt 1020 cgtccagcac ccaggccagc ctggagatcg actccctgtt tgagggcatc gacttctaca 1080 cgtccatcac cagggcgagg ttcgaggagc tgtgctccga cctgttccga agcaccctgg 1140 agcccgtgga gaaggctctg cgcgacgcca agctggacaa ggcccagatt cacgacctgg 1200 tcctggtcgg gggctccacc cgcatcccca aggtgcagaa gctgctgcag gacttcttca 1260 acgggcgcga cctgaacaag agcatcaacc ccgacgaggc tgtggcctac ggggcggcgg 1320 tgcaggcggc catcctgatg ggggacaagt ccgagaacgt gcaggacctg ctgctgctgg 1380 acgtggctcc cctgtcgctg gggctggaga cggccggagg cgtgatgact gccctgatca 1440 agcgcaactc caccatcccc accaagcaga cgcagatctt caccacctac tccgacaacc 1500 aacccggggt gctgatccag gtgtacgagg gcgagagggc catgacgaaa gacaacaatc 1560 tgttggggcg cttcgagctg agcggcatcc ctccggcccc caggggcgtg ccccagatcg 1620 aggtgacctt cgacatcgat gccaacggca tcctgaacgt cacggccacg gacaagagca 1680 ccggcaaggc caacaagatc accatcacca acgacaaggg ccgcctgagc aaggaggaga 1740 tcgagcgcat ggtgcaggag gcggagaagt acaaagcgga ggacgaggtg cagcgcgaga 1800 gggtgtcagc caagaacgcc ctggagtcct acgccttcaa catgaagagc gccgtggagg 1860 atgaggggct caagggcaag atcagcgagg ccgacaagaa gaaggtgctg gacaagtgtc 1920 aagaggtcat ctcgtggctg gacgccaaca ccttggccga gaaggacgag tttgagcaca 1980 agaggaagga gctggagcag gtgtgtaacc ccatcatcag cggactgtac cagggtgccg 2040 gtggtcccgg gcctgggggc ttcggggctc agggtcccaa gggagggtct gggtcaggcc 2100 ccaccattga ggaggtagat taggggcctt tccaagattg ctgtttttgt tttggagctt 2160 caagactttg catttcctag tatttctgtt tgtcagttct caatttcctg tgtttgcaat 2220 gttgaaattt tttggtgaag tactgaactt gctttttttc cggtttctac atgcagagat 2280 gaatttatac tgccatctta cgactatttc ttctttttaa tacacttaac tcaggccatt 2340 ttttaagttg gttacttcaa agtaaataaa ctttaaaatt caa 2383 <210> 103 <211> 3283 <212> DNA <213> human <400> 103 gcgtgtcggg cgcggaaggg ggaggcggcc cggggcgccc gcgagtgagg cgcggggcgg 60 cgaagggagc gcgggtggcg gcacttgctg ccgcggcctt ggatgggctg ggcccccctc 120 gccgctccgc ctcctccaca cgcgcggcgg ccgcggcgag ggggacgcgc cgcccggggc 180 ccggcacctt cgggaacccc ccggcccgga gcctgcggcc tgcgccgcct cggccgccgg 240 gagccccgtg gagcccccgc cgccgcgccg ccccgcggac cggacgctga gggcactcgg 300 ggcggggcgc gcgctcgggc agacgtttgc ggggaggggg gcgcctgccg ggccccggcg 360 accaccttgg gggtcgcggg ccggctcggg gggcgcccag tgcgggccct cgcgggcgcc 420 gggcagcgac cagccctgag cggagctgtt ggccgcggcg ggaggcctcc cggacgcccc 480 cagccccccg aacgctcgcc cgggccggcg ggagtcggcg ccccccggga ggtccgctcg 540 gtcgtccgcg gcggagcgtt tgctcctggg acaggcggtg ggaccggggc gtcgccggag 600 acgcccccag cgaagttggg ctctccaggt gtgggggtcc cggggggtag cgacgtcgcg 660 gacccggcct gtgggatggg cggcccggag aagactgcgc tcggccgtgt tcatacttgt 720 ccgtgggcct gaggtccccg gaggatgacc tagcactgaa aagccccggc cggcctcccc 780 agggtccccg aggacgaagt tgaccctgac cgggccgtct cccagttctg aggcccgggt 840 cccactggaa ctcgcgtctg agccgccgtc ccggaccccc ggtgcccgcc ggtccgcaga 900 ccctgcaccg ggcttggact cgcagccggg actgacgtgt agaacaatcg tttctgttgg 960 aagaagggtt tttcccttcc ttttggggtt tttgttgcct tttttttttc ttttttcttt 1020 gtaaaatttt ggagaaggga agtcggaaca caaggaagga ccgctcaccc gcggactcag 1080 ggctggcggc gggactccag gaccctgggt ccagcatgga ggtggtggac ccgcagcagc 1140 tgggcatgtt cacggagggc gagctgatgt cggtgggtat ggacacgttc atccaccgca 1200 tcgactccac cgaggtcatc taccagccgc gccgcaagcg ggccaagctc atcggcaagt 1260 acctgatggg ggacctgctg ggggaaggct cttacggcaa ggtgaaggag gtgctggact 1320 cggagacgct gtgcaggagg gccgtcaaga tcctcaagaa gaagaagttg cgaaggatcc 1380 ccaacgggga ggccaacgtg aagaaggaaa ttcaactact gaggaggtta cggcacaaaa 1440 atgtcatcca gctggtggat gtgttataca acgaagagaa gcagaaaatg tatatggtga 1500 tggagtactg cgtgtgtggc atgcaggaaa tgctggacag cgtgccggag aagcgtttcc 1560 cagtgtgcca ggcccacggg tacttctgtc agctgattga cggcctggag tacctgcata 1620 gccagggcat tgtgcacaag gacatcaagc cggggaacct gctgctcacc accggtggca 1680 ccctcaaaat ctccgacctg ggcgtggccg aggcactgca cccgttcgcg gcggacgaca 1740 cctgccggac cagccagggc tccccggctt tccagccgcc cgagattgcc aacggcctgg 1800 acaccttctc cggcttcaag gtggacatct ggtcggctgg ggtcaccctc tacaacatca 1860 ccacgggtct gtaccccttc gaaggggaca acatctacaa gttgtttgag aacatcggga 1920 aggggagcta cgccatcccg ggcgactgtg gccccccgct ctctgacctg ctgaaaggga 1980 tgcttgagta cgaaccggcc aagaggttct ccatccggca gatccggcag cacagctggt 2040 tccggaagaa acatcctccg gctgaagcac cagtgcccat cccaccgagc ccagacacca 2100 aggaccggtg gcgcagcatg actgtggtgc cgtacttgga ggacctgcac ggcgcggacg 2160 aggacgagga cctcttcgac atcgaggatg acatcatcta cactcaggac ttcacggtgc 2220 ccggacaggt cccagaagag gaggccagtc acaatggaca gcgccggggc ctccccaagg 2280 ccgtgtgtat gaacggcaca gaggcggcgc agctgagcac caaatccagg gcggagggcc 2340 gggcccccaa ccctgcccgc aaggcctgct ccgccagcag caagatccgc cggctgtcgg 2400 cctgcaagca gcagtgaggc tggccgcctg cagcccgtgt ccaggagccc cgccaggtgc 2460 ccgcgccagg ccctcagtct tcctgccggt tccgcccgcc ctcccggaga ggtggccgcc 2520 atgcttctgt gccgaccacg ccccaggacc tccggagcgc cctgcagggc cgggcagggg 2580 gacagcaggg accgggcgca gccctccccc ctcggccgcc cggcagtgca cgcggcttgt 2640 tgacttcgca gccccgggcg gagccttccc gggcgggcgt gggaggaggg aggcggcctc 2700 catgcacttt atgtggagac tactggcccc gcccgtggcc tcgtgctccg cagggcgccc 2760 agcgccgtcc ggcggccccg ccgcagacca gctggcgggt gtggagacca ggctcctgac 2820 cccgccatgc atgcagcgcc acctggaagc cgcgcggccg ctttggtttt ttgtttggtt 2880 ggttccattt tctttttttc tttttttttt taagaaaaaa taaaaggtgg atttgagctg 2940 tggctgtgag gggtgtttgg gagctgctgg gtggcagggg ggctgtgggg tcgggctcac 3000 gtcgcggccg cctttgcgct ctcgggtcac cctgctttgg cggcccggcc ggagggcagg 3060 accctcacct ctcccccaag gccactgcgc tcttgggacc ccagagaaaa cccggagcaa 3120 gcaggagtgt gcggtcaata tttatatcat ccagaaaaga aaaacacgag aaacgccatc 3180 gcgggatggt gcagacgcgg cggggactcg gagggtgccg tgcgggcgag gccgcccaaa 3240 tttggcaata aataaagctt gggaagcttg gacctgaaaa aaa 3283 <210> 104 <211> 2030 <212> DNA <213> human <400> 104 tggctggcaa tggccttgct gacctcgagc cgggcccacg tggggacctt tggagcacag 60 cctacgatcc tggtgcaagg ccggtggatg cagaggccag tccatatacc acccaggcct 120 gcgaggagcg tggtccccac ccatccagcc catatgtgca agtgcccttg acagagaggc 180 tggtcatatc catggtgacc atttatgggc cacaacaggt ccccatctgc gcagtgaacc 240 ctgtgctgag caccttgcag acgtgatctt gcttcgtcct gcagcactgt gcggggcagg 300 aaaatccaag aggaagaagg atctacggat atcctgcatg tccaagccac ccgcacccaa 360 ccccacaccc ccccggaacc tggactcccg gaccttcatc accattggag acagaaactt 420 tgaggtggag gctgatgact tggtgaccat ctcagaactg ggccgtggag cctatggggt 480 ggtagagaag gtgcggcacg cccagagcgg caccatcatg gccgtgaagc ggatccgggc 540 caccgtgaac tcacaggagc agaagcggct gctcatggac ctggacatca acatgcgcac 600 ggtcgactgt ttctacactg tcaccttcta cggggcacta ttcagagagg gagacgtgtg 660 gatctgcatg gagctcatgg acacatcctt ggacaagttc taccggaagg tgctggataa 720 aaacatgaca attccagagg acatccttgg ggagattgct gtgtctatcg tgcgggccct 780 ggagcatctg cacagcaagc tgtcggtgat ccacagagat gtgaagccct ccaatgtcct 840 tatcaacaag gagggccatg tgaagatgtg tgactttggc atcagtggct acttggtgga 900 ctctgtggcc aagacgatgg atgccggctg caagccctac atggcccctg agaggatcaa 960 cccagagctg aaccagaagg gctacaatgt caagtccgac gtctggagcc tgggcatcac 1020 catgattgag atggccatcc tgcggttccc ttacgagtcc tgggggaccc cgttccagca 1080 gctgaagcag gtggtggagg agccgtcccc ccagctccca gccgaccgtt tctcccccga 1140 gtttgtggac ttcactgctc agtgcctgag gaagaacccc gcagagcgta tgagctacct 1200 ggagctgatg gagcacccct tcttcacctt gcacaaaacc aagaagacgg acattgctgc 1260 cttcgtgaag aagatcctgg gagaagactc ataggggctg ggcctcggac cccactccgg 1320 ccctccagag ccccacagcc ccatctgcgg gggcagtgct cacccacacc ataagctact 1380 gccatcctgg cccagggcat ctgggaggaa ccgagggggc tgctcccacc tggctctgtg 1440 gcgagccatt tgtcccaagt gccaaagaag cagaccattg gggctcccag ccaggccctt 1500 gtcggcccca ccagtgcctc tccctgctgc tcctaggacc cgtctccagc tgctgagatc 1560 ctggactgag ggggcctgga tgccccctgt ggatgctgct gcccctgcac agcaggctgc 1620 cagtgcctgg gtggatgggc caccgccttg cccagcctgg atgccatcca agttgtatat 1680 ttttttaatc tctcgactga atggactttg cacactttgg cccagggtgg ccacacctct 1740 atcccggctt tggtgcgggg tacacaagag gggatgagtt gtgtgaatac cccaagactc 1800 ccatgaggga gatgccatga gccgcccaag gccttcccct ggcactggca aacagggcct 1860 ctgcggagca cactggctca cccagtcctg cccgccaccg ttatcggtgt cattcacctt 1920 tcgtgttttt tttaatttat cctctgttga ttttttcttt tgctttatgg gtttggcttg 1980 tttttcttgc atggtttgga gctgatcgct tctcccccac cccctagggg 2030 <210> 105 <211> 3583 <212> DNA <213> human <400> 105 aaaggcctgc agcaggacga ggacctgagc caggaatgca ggatggcggc ggtgaagaag 60 gaagggggtg ctctgagtga agccatgtcc ctggagggag atgaatggga actgagtaaa 120 gaaaatgtac aacctttaag gcaagggcgg atcatgtcca cgcttcaggg agcactggca 180 caagaatctg cctgtaacaa tactcttcag cagcagaaac gggcatttga atatgaaatt 240 cgattttaca ctggaaatga ccctctggat gtttgggata ggtatatcag ctggacagag 300 cagaactatc ctcaaggtgg gaaagagagt aatatgtcaa cgttattaga aagagctgta 360 gaagcactac aaggagaaaa acgatattat agtgatcctc gatttctcaa tctctggctt 420 aaattagggc gtttatgcaa tgagcctttg gatatgtaca gttacttgca caaccaaggg 480 attggtgttt cacttgctca gttctatatc tcatgggcag aagaatatga agctagagaa 540 aactttagga aagcagatgc gatatttcag gaagggattc aacagaaggc tgaaccacta 600 gaaagactac agtcccagca ccgacaattc caagctcgag tgtctcggca aactctgttg 660 gcacttgaga aagaagaaga ggaggaagtt tttgagtctt ctgtaccaca acgaagcaca 720 ctagctgaac taaagagcaa agggaaaaag acagcaagag ctccaatcat ccgtgtagga 780 ggtgctctca aggctccaag ccagaacaga ggactccaaa atccatttcc tcaacagatg 840 caaaataata gtagaattac tgtttttgat gaaaatgctg atgaggcttc tacagcagag 900 ttgtctaagc ctacagtcca gccatggata gcacccccca tgcccagggc caaagagaat 960 gagctgcaag caggcccttg gaacacaggc aggtccttgg aacacaggcc tcgtggcaat 1020 acagcttcac tgatagctgt acccgctgtg cttcccagtt tcactccata tgtggaagag 1080 actgcacaac agccagttat gacaccatgt aaaattgaac ctagtataaa ccacatccta 1140 agcaccagaa agcctggaaa ggaagaagga gatcctctac aaagggttca gagccatcag 1200 caagcgtctg aggagaagaa agagaagatg atgtattgta aggagaagat ttatgcagga 1260 gtaggggaat tctcctttga agaaattcgg gctgaagttt tccggaagaa attaaaagag 1320 caaagggaag ccgagctatt gaccagtgca gagaagagag cagaaatgca gaaacagatt 1380 gaagagatgg agaagaagct aaaagaaatc caaactactc agcaagaaag aacaggtgat 1440 cagcaagaag agacgatgcc tacaaaggag acaactaaac tgcaaattgc ttccgagtct 1500 cagaaaatac caggaatgac tctatccagt tctgtttgtc aagtaaactg ttgtgccaga 1560 gaaacttcac ttgcggagaa catttggcag gaacaacctc attctaaagg tcccagtgta 1620 cctttctcca tttttgatga gtttcttctt tcagaaaaga agaataaaag tcctcctgca 1680 gatcccccac gagttttagc tcaacgaaga ccccttgcag ttctcaaaac ctcagaaagc 1740 atcacctcaa atgaagatgt gtctccagat gtttgtgatg aatttacagg aattgaaccc 1800 ttgagcgagg atgccattat cacaggcttc agaaatgtaa caatttgtcc taacccagaa 1860 gacacttgtg actttgccag agcagctcgt tttgtatcca ctccttttca tgagataatg 1920 tccttgaagg atctcccttc tgatcctgag agactgttac cggaagaaga tctagatgta 1980 aagacctctg aggaccagca gacagcttgt ggcactatct acagtcagac tctcagcatc 2040 aagaagctga gcccaattat tgaagacagt cgtgaagcca cacactcctc tggcttctct 2100 ggttcttctg cctcggttgc aagcacctcc tccatcaaat gtcttcaaat tcctgagaaa 2160 ctagaactta ctaatgagac ttcagaaaac cctactcagt caccatggtg ttcacagtat 2220 cgcagacagc tactgaagtc cctaccagag ttaagtgcct ctgcagagtt gtgtatagaa 2280 gacagaccaa tgcctaagtt ggaaattgag aaggaaattg aattaggtaa tgaggattac 2340 tgcattaaac gagaatacct aatatgtgaa gattacaagt tattctgggt ggcgccaaga 2400 aactctgcag aattaacagt aataaaggta tcttctcaac ctgtcccatg ggacttttat 2460 atcaacctca agttaaagga acgtttaaat gaagattttg atcatttttg cagctgttat 2520 caatatcaag atggctgtat tgtttggcac caatatataa actgcttcac ccttcaggat 2580 cttctccaac acagtgaata tattacccat gaaataacag tgttgattat ttataacctt 2640 ttgacaatag tggagatgct acacaaagca gaaatagtcc atggtgactt gagtccaagg 2700 tgtctgattc tcagaaacag aatccacgat ccctatgatt gtaacaagaa caatcaagct 2760 ttgaagatag tggacttttc ctacagtgtt gaccttaggg tgcagctgga tgtttttacc 2820 ctcagcggct ttcggactgt acagatcctg gaaggacaaa agatcctggc taactgttct 2880 tctccctacc aggtagacct gtttggtata gcagatttag cacatttact attgttcaag 2940 gaacacctac aggtcttctg ggatgggtcc ttctggaaac ttagccaaaa tatttctgag 3000 ctaaaagatg gtgaattgtg gaataaattc tttgtgcgga ttctgaatgc caatgatgag 3060 gccacagtgt ctgttcttgg ggagcttgca gcagaaatga atggggtttt tgacactaca 3120 ttccaaagtc acctgaacaa agccttatgg aaggtaggga agttaactag tcctggggct 3180 ttgctctttc agtgagctag gcaatcaagt ctcacagatt gctgcctcag agcaatggtt 3240 gtattgtgga acactgaaac tgtatgtgct gtaatttaat ttaggacaca tttagatgca 3300 ctaccattgc tgttctactt tttggtacag gtatattttg acgtcactga tattttttat 3360 acagtgatat acttactcat ggccttgtct aacttttgtg aagaactatt ttattctaaa 3420 cagactcatt acaaatggtt accttgttat ttaacccatt tgtctctact tttccctgta 3480 cttttcccat ttgtaatttg taaaatgttc tcttatgatc accatgtatt ttgtaaataa 3540 taaaatagta tctgttaaaa aaaaaaaaaa aaaaaaaaaa aaa 3583 <210> 106 <211> 1379 <212> DNA <213> human <400> 106 ctgttggagc cgctgtggtt gctgtccgcg gagtggaagc gcgtgctttt gtttgtgtcc 60 ctggccatgg cgctgcagct ctcccgggag cagggaatca ccctgcgcgg gagcgccgaa 120 atcgtggccg agttcttctc attcggcatc aacagcattt tatatcagcg tggcatatat 180 ccatctgaaa cctttactcg agtgcagaaa tacggactca ccttgcttgt aactactgat 240 cttgagctca taaaatacct aaataatgtg gtggaacaac tgaaagattg gttatacaag 300 tgttcagttc agaaactggt tgtagttatc tcaaatattg aaagtggtga ggtcctggaa 360 agatggcagt ttgatattga gtgtgacaag actgcaaaag atgacagtgc acccagagaa 420 aagtctcaga aagctatcca ggatgaaatc cgttcagtga tcagacagat cacagctacg 480 gtgacatttc tgccactgtt ggaagtttct tgttcatttg atctgctgat ttatacagac 540 aaagatttgg ttgtacctga aaaatgggaa gagtcgggac cacagtttat taccaattct 600 gaggaagtcc gccttcgttc atttactact acaatccaca aagtaaatag catggtggcc 660 tacaaaattc ctgtcaatga ctgaggatga catgaggaaa ataatgtaat tgtaattttg 720 aaatgtggtt ttcctgaaat caagtcatct atagttgata tgttttattt cattggttaa 780 tttttacatg gagaaaacca aaatgatact tactgaactg tgtgtaattg ttcctttatt 840 tttttggtac ctatttgact taccatggag ttaacatcat gaatttattg cacattgttc 900 aaaaggaacc aggaggtttt tttgtcaaca ttgtgatgta tattcctttg aagatagtaa 960 ctgtagatgg aaaaacttgt gctataaagc tagatgcttt cctaaatcag atgttttggt 1020 caagtagttt gactcagtat aggtagggag atatttaagt ataaaataca acaaaggaag 1080 tctaaatatt cagaatcttt gttaaggtcc tgaaagtaac tcataatcta taaacaatga 1140 aatattgctg tatagctcct tttgaccttc atttcatgta tagttttccc tattgaatca 1200 gtttccaatt atttgacttt aatttatgta acttgaacct atgaagcaat ggatatttgt 1260 actgtttaat gttctgtgat acagaactct taaaaatgtt ttttcatgtg ttttataaaa 1320 tcaagtttta agtgaaagtg aggaaataaa gttaagtttg ttttaaattt gtcttaaaa 1379 <210> 107 <211> 2984 <212> DNA <213> human <400> 107 ggaaattcaa acgtgtttgc ggaaaggagt ttgggttcca tcttttcatt tccccagcgc 60 agctttctgt agaaatggaa tccgaggatt taagtggcag agaattgaca attgattcca 120 taatgaacaa agtgagagac attaaaaata agtttaaaaa tgaagacctt actgatgaac 180 taagcttgaa taaaatttct gctgatacta cagataactc gggaactgtt aaccaaatta 240 tgatgatggc aaacaaccca gaggactggt tgagtttgtt gctcaaacta gagaaaaaca 300 gtgttccgct aagtgatgct cttttaaata aattgattgg tcgttacagt caagcaattg 360 aagcgcttcc cccagataaa tatggccaaa atgagagttt tgctagaatt caagtgagat 420 ttgctgaatt aaaagctatt caagagccag atgatgcacg tgactacttt caaatggcca 480 gagcaaactg caagaaattt gcttttgttc atatatcttt tgcacaattt gaactgtcac 540 aaggtaatgt caaaaaaagt aaacaacttc ttcaaaaagc tgtagaacgt ggagcagtac 600 cactagaaat gctggaaatt gccctgcgga atttaaacct ccaaaaaaag cagctgcttt 660 cagaggagga aaagaagaat ttatcagcat ctacggtatt aactgcccaa gaatcatttt 720 ccggttcact tgggcattta cagaatagga acaacagttg tgattccaga ggacagacta 780 ctaaagccag gtttttatat ggagagaaca tgccaccaca agatgcagaa ataggttacc 840 ggaattcatt gagacaaact aacaaaacta aacagtcatg cccatttgga agagtcccag 900 ttaaccttct aaatagccca gattgtgatg tgaagacaga tgattcagtt gtaccttgtt 960 ttatgaaaag acaaacctct agatcagaat gccgagattt ggttgtgcct ggatctaaac 1020 caagtggaaa tgattcctgt gaattaagaa atttaaagtc tgttcaaaat agtcatttca 1080 aggaacctct ggtgtcagat gaaaagagtt ctgaacttat tattactgat tcaataaccc 1140 tgaagaataa aacggaatca agtcttctag ctaaattaga agaaactaaa gagtatcaag 1200 aaccagaggt tccagagagt aaccagaaac agtggcaatc taagagaaag tcagagtgta 1260 ttaaccagaa tcctgctgca tcttcaaatc actggcagat tccggagtta gcccgaaaag 1320 ttaatacaga gcagaaacat accacttttg agcaacctgt cttttcagtt tcaaaacagt 1380 caccaccaat atcaacatct aaatggtttg acccaaaatc tatttgtaag acaccaagca 1440 gcaatacctt ggatgattac atgagctgtt ttagaactcc agttgtaaag aatgactttc 1500 cacctgcttg tcagttgtca acaccttatg gccaacctgc ctgtttccag cagcaacagc 1560 atcaaatact tgccactcca cttcaaaatt tacaggtttt agcatcttct tcagcaaatg 1620 aatgcatttc ggttaaagga agaatttatt ccattttaaa gcagatagga agtggaggtt 1680 caagcaaggt atttcaggtg ttaaatgaaa agaaacagat atatgctata aaatatgtga 1740 acttagaaga agcagataac caaactcttg atagttaccg gaacgaaata gcttatttga 1800 ataaactaca acaacacagt gataagatca tccgacttta tgattatgaa atcacggacc 1860 agtacatcta catggtaatg gagtgtggaa atattgatct taatagttgg cttaaaaaga 1920 aaaaatccat tgatccatgg gaacgcaaga gttactggaa aaatatgtta gaggcagttc 1980 acacaatcca tcaacatggc attgttcaca gtgatcttaa accagctaac tttctgatag 2040 ttgatggaat gctaaagcta attgattttg ggattgcaaa ccaaatgcaa ccagatacaa 2100 caagtgttgt taaagattct caggttggca cagttaatta tatgccacca gaagcaatca 2160 aagatatgtc ttcctccaga gagaatggga aatctaagtc aaagataagc cccaaaagtg 2220 atgtttggtc cttaggatgt attttgtact atatgactta cgggaaaaca ccatttcagc 2280 agataattaa tcagatttct aaattacatg ccataattga tcctaatcat gaaattgaat 2340 ttcccgatat tccagagaaa gatcttcaag atgtgttaaa gtgttgttta aaaagggacc 2400 caaaacagag gatatccatt cctgagctcc tggctcatcc ctatgttcaa attcaaactc 2460 atccagttaa ccaaatggcc aagggaacca ctgaagaaat gaaatatgtt ctgggccaac 2520 ttgttggtct gaattctcct aactccattt tgaaagctgc taaaacttta tatgaacact 2580 atagtggtgg tgaaagtcat aattcttcat cctccaagac ttttgaaaaa aaaaggggaa 2640 aaaaatgatt tgcagttatt cgtaatgtca aataccacct ataaaatata ttggactgtt 2700 atactcttga atccctgtgg aaatctacat ttgaagacaa catcactctg aagtgttatc 2760 agcaaaaaaa attcagtaga ttatctttaa aagaaaactg taaaaatagc aaccacttat 2820 ggtactgtat atattgtaga cttgttttct ctgttttatg ctcttgtgta atctacttga 2880 catcatttta ctcttggaat agtgggtgga tagcaagtat attctaaaaa actttgtaaa 2940 taaagttttg tggctaaaat gacactaaaa aaaaaaaaaa aaaa 2984 <210> 108 <211> 2166 <212> DNA <213> human <400> 108 gggggtcgct gtgcctgtcc ccgtgtgatc cgaaaagtgc tggcaaaatg cggctgctgc 60 ttcgcccggg ggggtaaggg ggtgccctgg tgaaattttc tatcctggcc aactcaaatg 120 aggatggcaa gagagtgagc ccttggagag agaggctgag catcatgaag agaagaccct 180 gggagttccc tggggaccct gagtctggag ctggggcaag gccccccagg gcccgtcagt 240 ggcctgagga gatggttgga tcattccaaa cattgtctca gtgtggaaac tgaggcagac 300 agtggtcagg caggaccata tgagaactgg atgttggagc cagctctagc cacaggagag 360 gagctgccgg aactgacctt gctgaccaca ctgttggagg gccctggaga taagacgcag 420 ccacctgaag aggagacttt gtcccaagcc cctgagagtg aggaggaaca gaagaagaag 480 gctctggaaa ggagtatgta tgtcctgagt gaactggtag aaacagagaa aatgtacgtg 540 gacgacttgg ggcagattgt ggagggttat atggccacca tggctgctca gggggtcccc 600 gagagtcttc gaggccgtga caggattgtg tttgggaata tccagcaaat ctatgagtgg 660 caccgagact atttcttgca agagctacaa cggtgtctga aagatcctga ttggctggct 720 cagctattca tcaaacacga gcgccggctg catatgtatg tggtgtactg tcagaataag 780 cccaagtcag agcatgtggt gtcagagttt ggggacagct actttgagga gctccggcag 840 cagctggggc accgcctgca gctgaacgac ctcctcatca aacctgtgca gcggatcatg 900 aaataccagc tgctgctcaa ggattttctc aagtattaca atagagctgg gatggatact 960 gcagacctag agcaagctgt ggaggtcatg tgctttgtgc ccaagcgctg caacgatatg 1020 atgacgctgg ggagattgcg gggatttgag ggcaaactga ctgctcaggg gaagctcttg 1080 ggccaggaca ctttctgggt caccgagcct gaggctggag ggctgctatc ttcccgaggt 1140 cgagagaggc gcgtcttcct ctttgagcaa atcatcatct tcagtgaagc cctgggagga 1200 ggagtgagag gtggaacaca gcctggatat gtatacaaga acagcattaa ggtgagctgc 1260 ctgggactgg aggggaacct ccaaggtgac ccttgccgct ttgcactgac ctccagaggg 1320 ccagagggtg ggatccagcg ctatgtcctg caggctgcag accctgctat cagtcaggcc 1380 tggatcaagc atgtggctca gatcttggag agccaacggg acttcctcaa cgcattgcag 1440 tcacccattg agtaccagag acgggagagc cagaccaaca gcctggggcg gccaagaggg 1500 cctggagtgg ggagccctgg aagaattcgg cttggagatc aggcccaggg cagcacacac 1560 acacccatca atggctctct cccctctctg ctgctgtcac ccaaagggga ggtggccaga 1620 gccctcttgc cactggataa acaggccctt ggtgacatcc cccaggctcc ccatgactct 1680 cctccagtct ctccaactcc aaaaacccct ccctgccaag ccagacttgc caagctggat 1740 gaagatgagc tgtaactggt gaaaaccatg ggggtggtgc tgactcagcc gcctattccc 1800 caaggagctt cagggcagtc cttctggcac tgctccagaa ttcctccttc ttggtgtgtc 1860 tggagggtgg gcaaggctgg gagggatatc aacttggagg agaacaccta gacccaagga 1920 cttttttctg cccaaggaac acagtttcct tcagctccca tccctatgca tgcatcatgg 1980 tccccccaaa aggaggatat gtgggtgggt gggagggctg gggcaggggc cagatagaaa 2040 ttattggttt tgttttttaa ttttgttttt cctgttttct gagaataaag gttttgttat 2100 atcaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 2160 aaaaaa 2166 <210> 109 <211> 3486 <212> DNA <213> human <400> 109 gagccgactg cggctgcgcg gggtattcga atcggcggcg gcttctagtt tgcggttcag 60 gtttggccgc tgccggccag cgtcctctgg ccatggacac cccggaaaat gtccttcaga 120 tgcttgaagc ccacatgcag agctacaagg gcaatgaccc tcttggtgaa tgggaaagat 180 acatacagtg ggtagaagag aattttcctg agaataaaga atacttgata actttactag 240 aacatttaat gaaggaattt ttagataaga agaaatacca caatgaccca agattcatca 300 gttattgttt aaaatttgct gagtacaaca gtgacctcca tcaatttttt gagtttctgt 360 acaaccatgg gattggaacc ctgtcatccc ctctgtacat tgcctgggcg gggcatctgg 420 aagcccaagg agagctgcag catgccagtg ctgtccttca gagaggaatt caaaaccagg 480 ctgaacccag agagttcctg caacaacaat acaggttatt tcagacacgc ctcactgaaa 540 cccatttgcc agctcaagct agaacctcag aacctctgca taatgttcag gttttaaatc 600 aaatgataac atcaaaatca aatccaggaa ataacatggc ctgcatttct aagaatcagg 660 gttcagagct ttctggagtg atatcttcag cttgtgataa agagtcaaat atggaacgaa 720 gagtgatcac gatttctaaa tcagaatatt ctgtgcactc atctttggca tccaaagttg 780 atgttgagca ggttgttatg tattgcaagg agaagcttat tcgtggggaa tcagaatttt 840 cctttgaaga attgagagcc cagaaataca atcaacggag aaagcatgag caatgggtaa 900 atgaagacag acattatatg aaaaggaaag aagcaaatgc ttttgaagaa cagctattaa 960 aacagaaaat ggatgaactt cataagaagt tgcatcaggt ggtggagaca tcccatgagg 1020 atctgcccgc ttcccaggaa aggtccgagg ttaatccagc acgtatgggg ccaagtgtag 1080 gctcccagca ggaactgaga gcgccatgtc ttccagtaac ctatcagcag acaccagtga 1140 acatggaaaa gaacccaaga gaggcacctc ctgttgttcc tcctttggca aatgctattt 1200 ctgcagcttt ggtgtcccca gccaccagcc agagcattgc tcctcctgtt cctttgaaag 1260 cccagacagt aacagactcc atgtttgcag tggccagcaa agatgctgga tgtgtgaata 1320 agagtactca tgaattcaag ccacagagtg gagcagagat caaagaaggg tgtgaaacac 1380 ataaggttgc caacacaagt tcttttcaca caactccaaa cacatcactg ggaatggttc 1440 aggcaacgcc atccaaagtg cagccatcac ccaccgtgca cacaaaagaa gcattaggtt 1500 tcatcatgaa tatgtttcag gctcctacac ttcctgatat ttctgatgac aaagatgaat 1560 ggcaatctct agatcaaaat gaagatgcat ttgaagccca gtttcaaaaa aatgtaaggt 1620 catctggggc ttggggagtc aataagatca tctcttcttt gtcatctgct tttcatgtgt 1680 ttgaagatgg aaacaaagaa aattatggat taccacagcc taaaaataaa cccacaggag 1740 ccaggacctt tggagaacgc tctgtcagca gacttccttc aaaaccaaag gaggaagtgc 1800 ctcatgctga agagtttttg gatgactcaa ctgtatgggg tattcgctgc aacaaaaccc 1860 tggcacccag tcctaagagc ccaggagact tcacatctgc tgcacaactt gcgtctacac 1920 cattccacaa gcttccagtg gagtcagtgc acattttaga agataaagaa aatgtggtag 1980 caaaacagtg tacccaggcg actttggatt cttgtgagga aaacatggtg gtgccttcaa 2040 gggatggaaa attcagtcca attcaagaga aaagcccaaa acaggccttg tcgtctcaca 2100 tgtattcagc atccttactt cgtctgagcc agcctgctgc aggtggggta cttacctgtg 2160 aggcagagtt gggcgttgag gcttgcagac tcacagacac tgacgctgcc attgcagaag 2220 atccaccaga tgctattgct gggctccaag cagaatggat gcagatgagt tcacttggga 2280 ctgttgatgc tccaaacttc attgttggga acccatggga tgataagctg attttcaaac 2340 ttttatctgg gctttctaaa ccagtgagtt cctatccaaa tacttttgaa tggcaatgta 2400 aacttccagc catcaagccc aagactgaat ttcaattggg ttctaagctg gtctatgtcc 2460 atcaccttct tggagaagga gcctttgccc aggtgtacga agctacccag ggagatctga 2520 atgatgctaa aaataaacag aaatttgttt taaaggtcca aaagcctgcc aacccctggg 2580 aattctacat tgggacccag ttgatggaaa gactaaagcc atctatgcag cacatgttta 2640 tgaagttcta ttctgcccac ttattccaga atggcagtgt attagtagga gagctctaca 2700 gctatggaac attattaaat gccattaacc tctataaaaa tacccctgaa aaagtgatgc 2760 ctcaaggtct tgtcatctct tttgctatga gaatgcttta catgattgag caagtgcatg 2820 actgtgaaat cattcatgga gacattaaac cagacaattt catacttgga aacggatttt 2880 tggaacagga tgatgaagat gatttatctg ctggcttggc actgattgac ctgggtcaga 2940 gtatagatat gaaacttttt ccaaaaggaa ctatattcac agcaaagtgt gaaacatctg 3000 gttttcagtg tgttgagatg ctcagcaaca aaccatggaa ctaccagatc gattactttg 3060 gggttgctgc aacagtatat tgcatgctct ttggcactta catgaaagtg aaaaatgaag 3120 gaggagagtg taagcctgaa ggtcttttta gaaggcttcc tcatttggat atgtggaatg 3180 aattttttca tgttatgttg aatattccag attgtcatca tcttccatct ttggatttgt 3240 taaggcaaaa gctgaagaaa gtatttcaac aacactatac taacaagatt agggccctac 3300 gtaataggct aattgtactg ctcttagaat gtaagcgttc acgaaaataa aatttggata 3360 tagacagtcc ttaaaaatca cactgtaaat atgaatctgc tcactttaaa cctgtttttt 3420 tttcatttat tgtttatgta aatgtttgtt aaaaataaat cccatggaat atttccatgt 3480 aaaaaa 3486 <210> 110 <211> 6641 <212> DNA <213> human <400> 110 ggttacattt tggatcctcg cggagtactg gtcaggcggt taagtcctgt acctaggaaa 60 gagggcgagc tctggggcgc tctccggtgt catgaggagc ttcaaaagag tcaactttgg 120 gactctgcta agcagccaga aggaggctga agagttgctg cccgccttga aggagttcct 180 gtccaaccct ccagctggtt ttcccagcag ccgatctgat gctgagagga gacaagcttg 240 tgatgccatc ctgagggctt gcaaccagca gctgactgct aagctagctt gccctaggca 300 tctggggagc ctgctggagc tggcagagct ggcctgtgat ggctacttag tgtctacccc 360 acagcgtcct cccctctacc tggaacgaat tctctttgtc ttactgcgga atgctgctgc 420 acaaggaagc ccagaggcca cactccgcct tgctcagccc ctccatgcct gcttggtgca 480 gtgctctcgc gaggctgctc cccaggacta tgaggccgtg gctcggggca gcttttctct 540 gctttggaag ggggcagaag ccctgttgga acggcgagct gcatttgcag ctcggctgaa 600 ggccttgagc ttcctagtac tcttggagga tgaaagtacc ccttgtgagg ttcctcactt 660 tgcttctcca acagcctgtc gagcggtagc tgcccatcag ctatttgatg ccagtggcca 720 tggtctaaat gaagcagatg ctgatttcct agatgacctg ctctccaggc acgtgatcag 780 agccttggtg ggtgagagag ggagctcttc tgggcttctt tctccccaga gggccctctg 840 cctcttggag ctcaccttgg aacactgccg tcgcttttgc tggagccgcc accatgacaa 900 agccatcagc gcagtggaga aggctcacag ttacctaagg aacaccaatc tagcccctag 960 ccttcagcta tgtcagctgg gggttaagct gctgcaggtt ggggaggaag gacctcaggc 1020 agtggccaag cttctgatca aggcatcagc tgtcctgagc aagagtatgg aggcaccatc 1080 acccccactt cgggcattgt atgagagctg ccagttcttc ctttcaggcc tggaacgagg 1140 caccaagagg cgctatagac ttgatgccat tctgagcctc tttgcttttc ttggagggta 1200 ctgctctctt ctgcagcagc tgcgggatga tggtgtgtat gggggctcct ccaagcaaca 1260 gcagtctttt cttcagatgt actttcaggg acttcacctc tacactgtgg tggtttatga 1320 ctttgcccaa ggctgtcaga tagttgattt ggctgacctg acccaactag tggacagttg 1380 taaatctacc gttgtctgga tgctggaggc cttagagggc ctgtcgggcc aagagctgac 1440 ggaccacatg gggatgaccg cttcttacac cagtaatttg gcctacagct tctatagtca 1500 caagctctat gccgaggcct gtgccatctc tgagccgctc tgtcagcacc tgggtttggt 1560 gaagccaggc acttatcccg aggtgcctcc tgagaagttg cacaggtgct tccggctaca 1620 agtagagagt ttgaagaaac tgggtaaaca ggcccagggc tgcaagatgg tgattttgtg 1680 gctggcagcc ctgcaaccct gtagccctga acacatggct gagccagtca ctttctgggt 1740 tcgggtcaag atggatgcgg ccagggctgg agacaaggag ctacagctaa agactctgcg 1800 agacagcctc agtggctggg acccggagac cctggccctc ctgctgaggg aggagctgca 1860 ggcctacaag gcggtgcggg ccgacactgg acaggaacgc ttcaacatca tctgtgacct 1920 cctggagctg agccccgagg agacaccagc cggggcctgg gcacgagcca cccacctggt 1980 agaactggct caggtgctct gctaccacga ctttacgcag cagaccaact gctctgctct 2040 ggatgctatc cgggaagccc tgcagcttct ggactctgtg aggcctgagg cccaggccag 2100 agatcagctt ctggacgata aagcacaggc cttgctgtgg ctttacatct gtactctgga 2160 agccaaaatg caggaaggta tcgagcggga tcggagagcc caggcccctg gtaacttgga 2220 ggaatttgaa gtcaatgacc tgaactatga agataaactc caggaagatc gtttcctata 2280 cagtaacatt gccttcaacc tggctgcaga tgctgctcag tccaaatgcc tggaccaagc 2340 cctggccctg tggaaggagc tgcttacaaa ggggcaggcc ccagctgtac ggtgtctcca 2400 gcagacagca gcctcactgc agatcctagc agccctctac cagctggtgg caaagcccat 2460 gcaggctctg gaggtcctcc tgctgctacg gattgtctct gagagactga aggaccactc 2520 gaaggcagct ggctcctcct gccacatcac ccagctcctc ctgaccctcg gctgtcccag 2580 ctatgcccag ttacacctgg aagaggcagc atcgagcctg aagcatctcg atcagactac 2640 tgacacatac ctgctccttt ccctgacctg tgatctgctt cgaagtcaac tctactggac 2700 tcaccagaag gtgaccaagg gtgtctctct gctgctgtct gtgcttcggg atcctgccct 2760 ccagaagtcc tccaaggctt ggtacttgct gcgtgtccag gtcctgcagc tggtggcagc 2820 ttaccttagc ctcccgtcaa acaacctctc acactccctg tgggagcagc tctgtgccca 2880 aggctggcag acacctgaga tagctctcat agactcccat aagctcctcc gaagcatcat 2940 cctcctgctg atgggcagtg acattctctc aactcagaaa gcagctgtgg agacatcgtt 3000 tttggactat ggtgaaaatc tggtacaaaa atggcaggtt ctttcagagg tgctgagctg 3060 ctcagagaag ctggtctgcc acctgggccg cctgggtagt gtgagtgaag ccaaggcctt 3120 ttgcttggag gccctaaaac ttacaacaaa gctgcagata ccacgccagt gtgccctgtt 3180 cctggtgctg aagggcgagc tggagctggc ccgcaatgac attgatctct gtcagtcgga 3240 cctgcagcag gttctgttct tgcttgagtc ttgcacagag tttggtgggg tgactcagca 3300 cctggactct gtgaagaagg tccacctgca gaaggggaag cagcaggccc aggtcccctg 3360 tcctccacag ctcccagagg aggagctctt cctaagaggc cctgctctag agctggtggc 3420 cactgtggcc aaggagcctg gccccatagc accttctaca aactcctccc cagtcttgaa 3480 aaccaagccc cagcccatac ccaacttcct gtcccattca cccacctgtg actgctcgct 3540 ctgcgccagc cctgtcctca cagcagtctg tctgcgctgg gtattggtca cggcaggggt 3600 gaggctggcc atgggccacc aagcccaggg tctggatctg ctgcaggtcg tgctgaaggg 3660 ctgtcctgaa gccgctgagc gcctcaccca agctctccaa gcttccctga atcataaaac 3720 acccccctcc ttggttccaa gcctcttgga tgagatcttg gctcaagcat acacactgtt 3780 ggcactggag ggcctgaacc agccatcaaa cgagagcctg cagaaggttc tacagtcagg 3840 gctgaagttt gtagcagcac ggatacccca cctagagccc tggcgagcca gcctgctctt 3900 gatttgggcc ctcacaaaac taggtggcct cagctgctgt actacccaac tttttgcaag 3960 ctcctggggc tggcagccac cattaataaa aagtgtccct ggctcagagc cctctaagac 4020 tcagggccaa aaacgttctg gacgagggcg ccaaaagtta gcctctgctc ccctgcgcct 4080 caataatacc tctcagaaag gtctggaagg tagaggactg ccctgcacac ctaaaccccc 4140 agaccggatc aggcaagctg gccctcatgt ccccttcacg gtgtttgagg aagtctgccc 4200 tacagagagc aagcctgaag taccccaggc ccccagggta caacagagag tccagacgcg 4260 cctcaaggtg aacttcagtg atgacagtga cttggaagac cctgtctcag ctgaggcctg 4320 gctggcagag gagcctaaga gacggggcac tgcttcccgg ggccgggggc gagcaaggaa 4380 gggcctgagc ctaaagacgg atgccgtggt tgccccaggt agtgcccctg ggaaccctgg 4440 cctgaatggc aggagccgga gggccaagaa ggtggcatca agacattgtg aggagcggcg 4500 tccccagagg gccagtgacc aggccaggcc tggccctgag atcatgagga ccatccctga 4560 ggaagaactg actgacaact ggagaaaaat gagctttgag atcctcaggg gctctgacgg 4620 ggaagactca gcctcaggtg ggaagactcc agctccgggc cctgaggcag cttctggaga 4680 atgggagctg ctgaggctgg attccagcaa gaagaagctg cccagcccat gcccagacaa 4740 ggagagtgac aaggaccttg gtcctcggct ccggctcccc tcagcccccg tagccactgg 4800 tctttctacc ctggactcca tctgtgactc cctgagtgtt gctttccggg gcattagtca 4860 ctgtcctcct agtgggctct atgcccacct ctgccgcttc ctggccttgt gcctgggcca 4920 ccgggatcct tatgccactg ctttccttgt caccgagtct gtctccatca cctgtcgcca 4980 ccagctgctc acccacctcc acagacagct cagcaaggcc cagaagcacc gaggatcact 5040 tgaaatagca gaccagctgc aggggctgag ccttcaggag atgcctggag atgtccccct 5100 ggcccgcatc cagcgcctct tttccttcag ggctttggaa tctggccact tcccccagcc 5160 tgaaaaggag agtttccagg agcgcctggc tctgatcccc agtggggtga ctgtgtgtgt 5220 gttggccctg gccaccctcc agcccggaac cgtgggcaac accctcctgc tgacccggct 5280 ggaaaaggac agtcccccag tcagtgtgca gattcccact ggccagaaca agcttcatct 5340 gcgttcagtc ctgaatgagt ttgatgccat ccagaaggca cagaaagaga acagcagctg 5400 tactgacaag cgagaatggt ggacagggcg gctggcactg gaccacagga tggaggttct 5460 catcgcttcc ctagagaagt ctgtgctggg ctgctggaag gggctgctgc tgccgtccag 5520 tgaggagccc ggccctgccc aggaggcctc ccgcctacag gagctgctac aggactgtgg 5580 ctggaaatat cctgaccgca ctctgctgaa aatcatgctc agtggtgccg gtgccctcac 5640 ccctcaggac attcaggccc tggcctacgg gctgtgccca acccagccag agcgagccca 5700 ggagctcctg aatgaggcag taggacgtct acagggcctg acagtaccaa gcaatagcca 5760 ccttgtcttg gtcctagaca aggacttgca gaagctgccg tgggaaagca tgcccagcct 5820 ccaagcactg cctgtcaccc ggctgccctc cttccgcttc ctactcagct actccatcat 5880 caaagagtat ggggcctcgc cagtgctgag tcaaggggtg gatccacgaa gtaccttcta 5940 tgtcctgaac cctcacaata acctgtcaag cacagaggag caatttcgag ccaatttcag 6000 cagtgaagct ggctggagag gagtggttgg ggaggtgcca agacctgaac aggtgcagga 6060 agccctgaca aagcatgatt tgtatatcta tgcagggcat ggggctggtg cccgcttcct 6120 tgatgggcag gctgtcctgc ggctgagctg tcgggcagtg gccctgctgt ttggctgtag 6180 cagtgcggcc ctggctgtgc gtggaaacct ggagggggct ggcatcgtgc tcaagtacat 6240 catggctggt tgccccttgt ttctgggtaa tctctgggat gtgactgacc gcgacattga 6300 ccgctacacg gaagctctgc tgcaaggctg gcttggagca ggcccagggg ccccccttct 6360 ctactatgta aaccaggccc gccaagctcc ccgactcaag tatcttattg gggctgcacc 6420 tatagcctat ggcttgcctg tctctctgcg gtaaccccat ggagctgtct tattgatgct 6480 agaagcctca taactgttct acctccaagg ttagatttaa tccttaggat aactctttta 6540 aagtgatttt ccccagtgtt ttatatgaaa catttccttt tgatttaacc tcagtataat 6600 aaagatacat catttaaacc ctgaaaaaaa aaaaaaaaaa a 6641 <210> 111 <211> 2080 <212> DNA <213> human <400> 111 gcggccgctg ggctcaggct gtccgctctg ggctccgcgg cctcggcccc gctgcactcc 60 acctccgccc cctcggactc cctcccctct gcttctactc ctcctgctcc agtgcggatc 120 gtttcgcaac tgcttgccac tcgtcccgtg cctggctgtt tttccatttc ccggccccct 180 cttcttgagt actttacccc ctgcatttgg ggacagggac tggaaaaggg gcgggtggag 240 cgtccagtgg agaagaagga agcgaggccc gcaggaggag gaggatcggc ggactgtggg 300 gaggagaccc cacgccaccc tttctggtca tctcccctcc cgccccgccc ctgcgcacac 360 tccctcgcgg gcgagctact ttcggaccag gaaagtaaga gcggccctgg gtgacagcgc 420 cgcggggcca gtcccggggt tagccgcgcg tctgctcgct tctggtccgt cgcgctccca 480 gccagggcac agcccggacc gaggatggct tcgaccacaa cctgcaccag gttcacggac 540 gagtatcagc ttttcgagga gcttggaaag ggggcattct cagtggtgag aagatgtatg 600 aaaattccta ctggacaaga atatgctgcc aaaattatca acaccaaaaa gctttctgct 660 agggatcatc agaaactaga aagagaagct agaatctgcc gtcttttgaa gcaccctaat 720 attgtgcgac ttcatgatag catatcagaa gagggctttc actacttggt gtttgattta 780 gttactggag gtgaactgtt tgaagacata gtggcaagag aatactacag tgaagctgat 840 gccagtcatt gtatacagca gattctagaa agtgttaatc attgtcacct aaatggcata 900 gttcacaggg acctgaagcc tgagaatttg cttttagcta gcaaatccaa gggagcagct 960 gtgaaattgg cagactttgg cttagccata gaagttcaag gggaccagca ggcgtggttt 1020 ggttttgctg gcacacctgg atatctttct ccagaagttt tacgtaaaga tccttatgga 1080 aagccagtgg atatgtgggc atgtggtgtc attctctata ttctacttgt ggggtatcca 1140 cccttctggg atgaagacca acacagactc tatcagcaga tcaaggctgg agcttatgat 1200 tttccatcac cagaatggga cacggtgact cctgaagcca aagacctcat caataaaatg 1260 cttactatca accctgccaa acgcatcaca gcctcagagg cactgaagca cccatggatc 1320 tgtcaacgtt ctactgttgc ttccatgatg cacagacagg agactgtaga ctgcttgaag 1380 aaatttaatg ctagaagaaa actaaagggt gccatcttga caactatgct ggctacaagg 1440 aatttctcag cagccaagag tttgttgaag aaaccagatg gagtaaagga gtcaactgag 1500 agttcaaata caacaattga ggatgaagat gtgaaagcac gaaagcaaga gattatcaaa 1560 gtcactgaac aactgatcga agctatcaac aatggggact ttgaagccta cacaaaaatc 1620 tgtgacccag gccttactgc ttttgaacct gaagctttgg gtaatttagt ggaagggatg 1680 gattttcacc gattctactt tgaaaatgct ttgtccaaaa gcaataaacc aatccacact 1740 attattctaa accctcatgt acatctggta ggggatgatg ccgcctgcat agcatatatt 1800 aggctcacac agtacatgga tggcagtgga atgccaaaga caatgcagtc agaagagact 1860 cgtgtgtggc accgccggga tggaaagtgg cagaatgttc attttcatcg ctcggggtca 1920 ccaacagtac ccatcaagcc accctgtatt ccaaatggga aagaaaactt ctcaggaggc 1980 acctctttgt ggcaaaacat ctaaggcctg aaaaccattc acatatgggt cttctaaatt 2040 tcaacagtgc cacttctgca ttctctgttc tcaaggcacc 2080 <210> 112 <211> 11611 <212> DNA <213> human <400> 112 ggcttcagcc ctgcagggaa agaaaagtgc aatgattctg gactgagacg cgcttgggca 60 gaggctatgt aatcgtgtct gtgttgagga cttcgcttcg aggagggaag aggagggatc 120 ggctcgctcc tccggcggcg gcggcggcgg cgactctgca ggcggagttt cgcggcggcg 180 gcaccagggt tacgccagcc ccgcggggag gtctctccat ccagcttctg cagcggcgaa 240 agccccagcg cccgagcgcc tgagccggcg gggagcaagt aaagctagac cgatctccgg 300 ggagccccgg agtaggcgag cggcggccgc cagctagttg agcgcacccc ccgcccgccc 360 cagcggcgcc gcggcgggcg gcgtccaggc ggcatggaga aggacggcct gtgccgcgct 420 gaccagcagt acgaatgcgt ggcggagatc ggggagggcg cctatgggaa ggtgttcaag 480 gcccgcgact tgaagaacgg aggccgtttc gtggcgttga agcgcgtgcg ggtgcagacc 540 ggcgaggagg gcatgccgct ctccaccatc cgcgaggtgg cggtgctgag gcacctggag 600 accttcgagc accccaacgt ggtcaggttg tttgatgtgt gcacagtgtc acgaacagac 660 agagaaacca aactaacttt agtgtttgaa catgtcgatc aagacttgac cacttacttg 720 gataaagttc cagagcctgg agtgcccact gaaaccataa aggatatgat gtttcagctt 780 ctccgaggtc tggactttct tcattcacac cgagtagtgc atcgcgatct aaaaccacag 840 aacattctgg tgaccagcag cggacaaata aaactcgctg acttcggcct tgcccgcatc 900 tatagtttcc agatggctct aacctcagtg gtcgtcacgc tgtggtacag agcacccgaa 960 gtcttgctcc agtccagcta cgccaccccc gtggatctct ggagtgttgg ctgcatattt 1020 gcagaaatgt ttcgtagaaa gcctcttttt cgtggaagtt cagatgttga tcaactagga 1080 aaaatcttgg acgtgattgg actcccagga gaagaagact ggcctagaga tgttgccctt 1140 cccaggcagg cttttcattc aaaatctgcc caaccaattg agaagtttgt aacagatatc 1200 gatgaactag gcaaagacct acttctgaag tgtttgacat ttaacccagc caaaagaata 1260 tctgcctaca gtgccctgtc tcacccatac ttccaggacc tggaaaggtg caaagaaaac 1320 ctggattccc acctgccgcc cagccagaac acctcggagc tgaatacagc ctgaggcctc 1380 agcagccgcc ttaagctgat cctgcggaga acacccttgg tggcttatgg gtccccctca 1440 gcaagcccta cagagctgtg gaggattgct atctggaggc cttccagctg ctgtcttctg 1500 gacaggctct gcttctccaa ggaaaccgcc tagtttactg ttttgaaatc aatgcaagag 1560 tgattgcagc tttatgttca tttgtttgtt tgtttgtctg tttgtttcaa gaacctggaa 1620 aaattccaga agaagagaag ctgctgacca attgtgctgc catttgattt ttctaacctt 1680 gaatgctgcc agtgtggagt gggtaatcca ggcacagctg agttatgatg taatctctct 1740 gcagctgccg ggcctgattt ggtacttttg agtgtgtgtg tgcatgtgtg tgtgtgtgtg 1800 tgtgtgtgtg tgtgtgtatg tgagagattc tgtgatcttt taaagtgtta ctttttgtaa 1860 acgacaagaa taattcaatt ttaaagactc aaggtggtca gtaaataaca ggcatttgtt 1920 cactgaaggt gattcaccaa aatagtcttc tcaaattaga aagttaaccc catgtcctca 1980 gcatttcttt tctggccaaa agcagtaaat ttgctagcag taaaagatga agttttatac 2040 acacagcaaa aaggagaaaa aattctagta tattttaaga gatgtgcatg cattctattt 2100 agtcttcaga atgctgaatt tacttgttgt aagtctattt taaccttctg tatgacatca 2160 tgctttatca tttcttttgg aaaatagcct gtaagctttt tattacttgc tataggttta 2220 gggagtgtac ctcagataga ttttaaaaaa aagaatagaa agcctttatt tcctggtttg 2280 aaattccttt cttccctttt tttgttgttg ttattgttgt ttgttgttgt tattttgttt 2340 ttgtttttag gaatttgtca gaaactcttt cctgttttgg tttggagagt agttctctct 2400 aactagagac aggagtggcc ttgaaatttt cctcatctat tacactgtac tttctgccac 2460 acactgcctt gttggcaaag tatccatctt gtctatctcc cggcacttct gaaatatatt 2520 gctaccattg tataactaat aacagattgc ttaagctgtt cccatgcacc acctgtttgc 2580 ttgctttcaa tgaacctttc ataaattcgc agtctcagct tatggtttat ggcctcgatt 2640 ctgcaaacct aacagggtca catatgttct ctaatgcagt ccttctacct ggtgtttact 2700 tttgctaccc aaataatgag taggatcttg ttttcgtata cccccaccac tcccattgct 2760 accaactgtc accttgtgca ctcctttttt atagaagata ttttcagtgt ctttacctga 2820 gggtatgtct ttagctatgt tttagggcca tacatttact ctatcaaatg atcttttctc 2880 catcccccag gctgtgctta tttctagtgc cttgtgctca ctcctgctct ctacagagcc 2940 agcctggcct gggcattgta aacagctttt cctttttctc ttactgtttt ctctacagtc 3000 ctttatattt cataccatct ctgccttata agtggtttag tgctcagttg gctctagtaa 3060 ccagaggaca cagaaagtat cttttggaaa gtttagccac ctgtgctttc tgactcagag 3120 tgcatgcaac agttagatca tgcaacagtt agattatgtt tagggttagg attttcaaag 3180 aatggaggtt gctgcactca gaaaataatt cagatcatgt ttatgcatta ttaagttgta 3240 ctgaattctt tgcagcttaa tgtgatatat gactatcttg aacaagagaa aaaactagga 3300 gatgtttctc ctgaagagct tttggggttg ggaactattc ttttttaatt gctgtactac 3360 ttaacattgt tctaattcag tagcttgagg aacaggaaca ttgttttcta gagcaagata 3420 ataaaggaga tgggccatac aaatgttttc tactttcgtt gtgacaacat tgattaggtg 3480 ttgtcagtac tataaatgct tgagatataa tgaatccaca gcattcaagg tcaggtctac 3540 tcaaagtctc acatggaaaa gtgagttctg cctttccttt gatcgagggt caaaatacaa 3600 agacattttt gctagggcct acaaattgaa tttaaaaact cactgcactg attcatctga 3660 gctttttggt tagtattcat ggctagagtg aacatagctt tagtttttgc tgttgtaaaa 3720 gtgttttcat aagttcactc aagaaaaatg cagctgttct gaactggaat ttttcagcat 3780 tctttagaat tttaaatgag tagagagctc aacttttatt cctagcatct gcttttgact 3840 catttctagg cagtgcttat gaagaaaaat taaagcacaa acattctggc attcaatcgt 3900 tggcagatta tcttctgatg acacagaatg aaagggcatc tcagcctctc tgaactttgt 3960 aaaaatctgt ccccagttct tccatcggtg tagttgttgc atttgagtga atactctctt 4020 gatttatgta ttttatgtcc agattcgcca tttctgaaat ccagatccaa cacaagcagt 4080 cttgccgtta gggcattttg aagcagatag tagagtaaga acttagtgac tacagcttat 4140 tcttctgtaa catatggttt caaacatctt tgccaaaagc taagcagtgg tgaactgaaa 4200 agggcatatt gccccaaggt tacactgaag cagctcatag caagttaaaa tattgtgaca 4260 gatttgaaat catgtttgaa tttcatagta ggaccagtac aagaatgtcc ctgctagttt 4320 ctgtttgatg tttggttctg gcggctcagg cattttggga actgttgcac agggtgcagt 4380 caaaacaacc tacatataaa aattacataa aagaaccttg tccatttagc tttcataaga 4440 aatcccatgg caaagagtaa taaaaaggac ctaatcttaa aaatacaatt tctaagcact 4500 tgtaagaacc cagtgggttg gagcctccca ctttgtccct cctttgaagt ggatgggaac 4560 tcaaggtgca aagaacctgt tttggaagaa agcttggggc catttcagcc ccctgtattc 4620 tcatgatttt ctctcaggaa gcacacactg tgaatggcag acttttcatt tagccccagg 4680 tgacttacta aaaatagttg aaaattattc acctaagaat agaatctcag cattgtgtta 4740 aataaaaatg aaagctttag aaggcatgag atgttcctat cttaaataaa gcatgtttct 4800 tttctataga gaaatgtata gtttgactct ccagaatgta ctatccatct tgatgagaaa 4860 actcttaaat agtaccaaac attttgaact ttaaattatg tatttaaagt gagtgtttaa 4920 gaaactgtag ctgcttcttt tacaagtggt gcctattaaa gtcagtaatg gccattattg 4980 ttccattgtg gaaattaaat tatgtaagct tcctaatatc ataaacatat taaaattctt 5040 ctaaaatatt gcttttcttt taagtgacaa tttgactatt cttatgataa gcacatgaga 5100 gtgtcttaca ttttccaaaa gcaggcttta attgcatagt tgagtctagg aaaaaataat 5160 gttaaaagtg aatatgccac cataattact taattatgtt agtatagaaa ctacagaata 5220 tttaccctgg aaagaaaata ttggaatgtt attataaact cttagatatt tatataattc 5280 aaaagaatgc atgtttcaca ttgtgacaga taaagatgta tgatttctaa ggctttaaaa 5340 attattcata aaacagtggg caatagataa aggaaattct ggagaaaatg aaggtattta 5400 aagggtagtt tcaaagctat atatattttg aaggatatat tctttatgaa caaatatatt 5460 gtaaaaattt atactaaggt catctggtaa ctgtgggatt aatatggtcg aaaacaaatg 5520 ttatggagaa gctgtcccaa gcaaactaaa ttacctgtac ttttttccca tttcaaggga 5580 agaggcaacc acatgaagca atacttctta cacatgccta agaacgttca ttgaaaaaat 5640 aaatttttaa aaggcatgtg tttcctatgc caccaatact tttgaaaaat tgtgaacctt 5700 acccaaaacc atttatcatg tccattaagt atatttgggt atataattag gaagatattt 5760 acatgttcca tctccacagt ggaaaaactt attgaggcta ccaaagtgtg ccaagaaatg 5820 taagtcctta gagtaattag aaatgctgtt ttcctcaaaa gcatgagaaa ctagcatttt 5880 catttcttat ttactccctt tctatatcaa tgcaattcac aacccaattt taatacatcc 5940 ctatatctca agcatttcta tcttgtactt tttcagaaaa taaaccaaaa ataatccttt 6000 ggtctctcta tcttctgacc tttgtaagca acagaaatgt aaaaacagaa ggggtccaat 6060 ttttacacgt ttttttctca agtagccttt ctggggattt ttattttctt aatgaagtgc 6120 caatcagctt ttcaaaatgt tttctatttc tcagcatttc caggaagtga taacgtttag 6180 ctaaatgagt agaagtggac ttccttcaac atattgttac cttgtctagc cttaggaaga 6240 aaacaagagc cacctgaaaa taaatacagg ctcttttcga gcatctgctg aaatactgtt 6300 acagcaattt gaagttgatg tggtaggaaa ggaaggtgac ttttcttgca aaagtctttc 6360 taaacattca cactgtccta agagatgagc tttcttgttt tattccggta tattccacaa 6420 ggtggcactt ttagagaaaa acaaatctga tgaagactaa agaggtactt ctaaaagaga 6480 tttcattcta actttatttt tctgcgcata tttaactctt tcctagcact tgttttttgg 6540 gatgattaat agtctctata atgttctgta acttcaatat tttacttgtt acctaggttc 6600 tgaacaattg tctgcaaata aattgttctt aaggatggat aatacaccca ttttgatcat 6660 ttaagtaaag aaagcctagt cattcattca gtcaagaaaa aatttttgaa gtacccagtt 6720 accttacttt tctagattaa aacaggctta gttactaaaa aggcagtcct catctgtgaa 6780 caggatagtt tcgttagaag tataaaactc ctttagtggc cccagttaaa acacacatac 6840 cctctctgct gctttcaaat tccctagcat ggtggccttt caacattgat taaattttaa 6900 aatcctaatt taaagatcag gtgagcaaaa tgagtagcac atcagtaatt cagtagacaa 6960 aacttttgtc tgaaaaattg ctgtattgaa acagagccct aaaataccaa aagaccaggt 7020 aattttaaca tttgtggaat cacaaatgta aattcataag aagctctaat taaaaaaaaa 7080 aagtctgaag tatatgagca taacaactta ggagtgtgtc tacatactta acttttgaag 7140 ttttttggca actttatata ctttttttaa atttacaagt ctacttaaag acttcttata 7200 ccccaaatga ttaagttaat tttagaggtc acctttctca cagcagtgtc acttgaaatt 7260 tagtagggaa ggatattgca gtatttttca gtttccttag cacagcacca cagaaagcag 7320 cttattcctt ttgagtggca gacactcgac ggtgcctgcc caactttcct cctgagtggc 7380 aagcagatga gtctcagtaa ttcatactga accaaaatgc cacatacact aggggcagtc 7440 agaaactggc tgagaaatcc cccgcctcat tcgcccctct gctcccagga actagagtcc 7500 agttaaagcc cctatgcgaa aggccgaatt ccaccccagg gtttgttata acagtggcca 7560 gtctgaaccc catttgctcg tgctcaaaac ttgattccca cttgaaagcc ttccgggcgc 7620 gctgcctcgt tgccccgccc ctttggcagg agagaggcag tgggcgaggc cgggctgggg 7680 ccccgcctcc cactcacctg ccggtgcctg aaattatgtg cggccccgcg ggctgctttc 7740 cgaggtcaga gtgccctgct gctgtctcag aggcatctgt tctgcaaatc ttaggaagaa 7800 aaatgtccct agtagcaaac gggtgtcttc tgtgcataaa taagtacaac acaattctcc 7860 gaaagttcgg gtaaaaagag atgcggtagc agctgccctg tgtgaagctg tctaccccgc 7920 atctctcagg cgctaagctc agtttttgtt tttgtttttg tttttttaaa gaaaagatgt 7980 ataattgcag gaattttttt ttattttttt attttccatc attctatata tgtgatggtg 8040 aaagatatgc ctggaaaagt tttgttttga aaagtttatt ttctgcttcg tcttcagttg 8100 gcaaaagctc tcaattcttt agcttccagt ttcttttctc tctttttctt tgttaggtaa 8160 ttaaaggtat gtaaacaaat tatctcatgt agcaggggat tttcatgttg agaggaatct 8220 tccgtgtgag ttgtttggtc acacaaataa ccctttctca attttaggag tttggattgt 8280 caaatgtagg tttttctcaa agggggcata taactacata ttgactgcca agaactatga 8340 ctgtagcact aatcagcaca catagagcca cacaattatt taatttctaa ctctctgtgg 8400 tccctagaaa aattccgttg atgtgcttag gttaaagttc tgaagatacc cgttgtaccc 8460 ttacttgaaa gtttctaatc ttaagtttta tgaaatgcaa taatatgtat cagctagcaa 8520 tatttctgtg atcaccaaca actctcagtt tgatcttaaa gtctgaataa taaaacaaat 8580 cccagcagta atacatttct taaacctcac agtgcatgat atatcttttc attctgatcc 8640 tgtgtttgca aaaatataca catgtatatc atagttcctc actttttatt catttgtttt 8700 cctattacct gtagtaaata tattagttag tacatggaat ttatagcatc agctaccccc 8760 aggaacagca cctgacaggc gggggatttt ttttcaagtt gttctacatt tgcataaatt 8820 atttctatta ttattcatgt atgttattta tttctgaatc acactagtcc tgtgaaagta 8880 caactgaagg cagaaagtgt taggattttg catctaatgt tcattatcat ggtattgatg 8940 gacctaagaa aataaaaatt agactaagcc cccaaataag ctgcatgcat ttgtaacatg 9000 attagtagat ttgaatatat agatgtagta ttttgggtat ctaggtgttt tatcattatg 9060 taaaggaatt aaagtaaagg actttgtagt tgtttttatt aaatatgcat atagtagagt 9120 gcaaaaatat agcaaaaata aaaactaaag gtagaaaagc attttagata tgccttaatt 9180 tagaaactgt gccaggtggc cctcggaata gatgccaggc agagaccagt gcctgggtgg 9240 tgcctcctct tgtctgccct catgaagaag cttccctcac gtgatgtagt gccctcgtag 9300 gtgtcatgtg gagtagtggg aacaggcagt actgttgaga ggagagcagt gtgagagttt 9360 ttctgtagaa gcagaactgt cagcttgtgc cttgaggctt ccagaacgtg tcagatggag 9420 aagtccaagt ttccatgctt caggcaactt agctgtgtac agaagcaatc cagtgtggta 9480 ataaaaagca aggattgcct gtataattta ttataaaata aaagggattt taacaaccaa 9540 caattcccaa cacctcaaaa gcttgttgca ttttttggta tttgaggttt ttatctgaag 9600 gttaaagggc aagtgtttgg tatagaagag cagtatgtgt taagaaaaga aaaatattgg 9660 ttcacgtaga gtgcaaatta gaactagaaa gttttatacg attatcattt tgagatgtgt 9720 taaagtaggt tttcactgta aaatgtatta gtgtttctgc attgccatag ggcctggtta 9780 aaactttctc ttaggtttca ggaagactgt cacatacagt aagctttttt ccttctgact 9840 tataatagaa aatgttttga aagtaaaaaa aaaaaatcta atttggaaat ttgacttgtt 9900 agtttctgtg tttgaaatca tggttctaga aatgtagaaa ttgtgtatat cagatactca 9960 tctaggctgt gtgaaccagc ccaagatgac caacatcccc acacctctac atctctgtcc 10020 cctgtatctc ttcctttcta ccactaaagt gttccctgct accatcctgg cttgtccaca 10080 tggtgctctc catcttcctc cacatcatgg accacaggtg tgcctgtcta ggcctggcca 10140 ccactcccaa cttgacctag ccacattcat ctagagatgg ttcctgatgc tgggcacaga 10200 ctgtgctcat ggcacccatt agaaatgcct ctagcatctt tgtatgcatc ttgattttta 10260 aaccaagtca ttgtacagag cattcagttt tggctgtggt accaagagaa aaactaatca 10320 agaatataaa ccacattcca ggctgctgtt ttctctccat ctacaggcca cacttttact 10380 gtatttcttc atacttgaaa ttcattctgc tattttcata tcagggtaca gacttataag 10440 ggtgcatgtt ccttaaaggt gcataattat tcttattccg tttgcttata ttgctacaga 10500 atgctctgtt ttggtgcttt gagttctgca gacccaagaa gcagtgtgga aattcactgc 10560 ctgggacaca gtcttataag aatgttggca ggtgactttg tatcagatgt tgcttctctt 10620 ttctctgtac acagattgag agttaccaca gtggcctgtc gggtccaccc tgtgggtgca 10680 gcacagctct ctgaaagcaa gaaccttcct acctattcta acgtttttgc cctctaagaa 10740 aaatggcctc aggtatggta tagacatagc aagaggggaa gggctgtctc actctagcaa 10800 ccatccctcc attacacaca gaaagccctc ttgaagcaaa agaagaagaa agaaagaaag 10860 cttatctcta aggctactgt cttcagaatg ctctgagctg aatgctcttg ctcctttccc 10920 aagaggcaga tgaaaatata gccagtttat ctataccctt cctatctgag gaggagaata 10980 gaaaagtagg gtaaatatgt aacgtaaaat atgtcattca aggaccacca aaactttaag 11040 taccctatca ttaaaaatct ggttttaaaa gtagctcaag taagggatgc tttgtgaccc 11100 agggtttctg aagtcagata gccattctta cctgcccctt actctgactt attgggaaag 11160 ggagaactgc agtggtgttt ctgttgcagt ggcaaaggta acatgtcaga aaattcagag 11220 ggttgcatac caataatcct ttggaaactg gatgtcttac tgggtgctag aatgaaaatg 11280 taggtattta ttgtcagatg atgaagttca ttgttttttt caaaattggt gttgaaatat 11340 cactgtccaa tgtgttcact tatgtgaaag ctaaattgaa tgaggcaaaa agagcaaata 11400 gtttgtatat ttgtaatacc ttttgtattt cttacaataa aaatattggt agcaaataaa 11460 aataataaaa acaataactt taaactgctt tctggagatg aattactctc ctggctattt 11520 tcttttttac tttaatgtaa aatgagtata actgtagtga gtaaaattca ttaaattcca 11580 agttttagca gaaaaaaaaa aaaaaaaaaa a 11611 <210> 113 <211> 3317 <212> DNA <213> human <400> 113 agttctcgcg ggacaccgac ggggagcgga agccaggagg tattgctgct tcggcgaccg 60 ggcggcggca gcggcggcgg cggctgtggc agagtctgtg cctgtggcgg tgacggcggc 120 gggagcaagc gctgccctcg cagagcagcc ttggggtcgc cggccgctcg cagcgttgtg 180 gaggggcggg ccggacgctg agcggagcag ctgcgccacg ggtggcattg tgtgtcccag 240 agtgccggag cgagtcccag aagagaggcg aggctaagcc cagagcgctg ggttgcttca 300 gcagggaaga ctcccttccc cctgcttcag gctgctgagc actgagcagc gctcagaatg 360 gaagccatcg ccaaatatga cttcaaagct actgcagacg acgagctgag cttcaaaagg 420 ggggacatcc tcaaggtttt gaacgaagaa tgtgatcaga actggtacaa ggcagagctt 480 aatggaaaag acggcttcat tcccaagaac tacatagaaa tgaaaccaca tccgtggttt 540 tttggcaaaa tccccagagc caaggcagaa gaaatgctta gcaaacagcg gcacgatggg 600 gcctttctta tccgagagag tgagagcgct cctggggact tctccctctc tgtcaagttt 660 ggaaacgatg tgcagcactt caaggtgctc cgagatggag ccgggaagta cttcctctgg 720 gtggtgaagt tcaattcttt gaatgagctg gtggattatc acagatctac atctgtctcc 780 agaaaccagc agatattcct gcgggacata gaacaggtgc cacagcagcc gacatacgtc 840 caggccctct ttgactttga tccccaggag gatggagagc tgggcttccg ccggggagat 900 tttatccatg tcatggataa ctcagacccc aactggtgga aaggagcttg ccacgggcag 960 accggcatgt ttccccgcaa ttatgtcacc cccgtgaacc ggaacgtcta agagtcaaga 1020 agcaattatt taaagaaagt gaaaaatgta aaacacatac aaaagaatta aacccacaag 1080 ctgcctctga cagcagcctg tgagggagtg cagaacacct ggccgggtca ccctgtgacc 1140 ctctcacttt ggttggaact ttagggggtg ggagggggcg ttggatttaa aaatgccaaa 1200 acttacctat aaattaagaa gagtttttat tacaaatttt cactgctgct cctctttccc 1260 ctcctttgtc ttttttttca tccttttttc tcttctgtcc atcagtgcat gacgtttaag 1320 gccacgtata gtcctagctg acgccaataa taaaaaacaa gaaaccaagt gggctggtat 1380 tctctctatg caaaatgtct gttttagttg gaatgactga aagaagaaca gctgttcctg 1440 tgttcttcgt atatacacac aaaaaggagc gggcagggcc gctcgatgcc tttgctgttt 1500 agcttcctcc agaggagggg acttgtagga atctgccttc cagcccagac ccccagtgta 1560 ttttgtccaa gttcacagta gagtagggta gaaggaaagc atgtctctgc ttccatggct 1620 tcctgagaaa gcccacctgg gctgggcgcg gtggctcacg cctgtaatcc cagcactttg 1680 ggaggccaag gtgggcggat cacaaggtca ggagttcgag accaacctag ccaacatggt 1740 gaaaccccgt ctctactaaa aataagaaat tagccgggtg tggcacgcac ctgtagtccc 1800 agctacttgg gagcctgagg caggagaatc gcttgaacct gggaagtgga ggttgagtga 1860 gccgggaccg tgccattgta ctccagcctg ggtgacagag cgagattccg tctcaaaaaa 1920 aaaaaaaaaa agcccacctg aaagcctgtc tctttccact ttgttggccc ttccagtggg 1980 attatcgagc atgttgtttt ttcatagtgc ctttttcctt atttcaaggg ttgcttctga 2040 gtggtgtttt tttttttttt ttaatttgtt ttgttttaaa ataagttaaa ggcagtccag 2100 agcttttcag ccaatttgtc tcctactctg tgtaaatatt tttccctccg ggcaggggag 2160 ccagggtaga gcaaaggaga caaagcagga gtggaaggtg aggcgttctc ctgcttgtac 2220 taagccagga ggctttaagc tccagcttta agggttgtga gccccttggg ggttcaggga 2280 actgcttgcc cagggtgcag tgtgagtgtg atgggccacc ggggcaagag ggaaggtgac 2340 cgcccagctc tcccacatcc cactggatct ggcttacagg ggggtcggaa gcctgtcctc 2400 accgtctcgg gggttgtggc ccccgccccc tccctatatg cacccctgga accagcaagt 2460 cccagacaag gagagcggag gaggaagtca tgggaacgca gcctccagtt gtagcaggtt 2520 tcactattcc tatgctgggg tacacagtga gagtactcac ttttcacttg tcttgctctt 2580 agattgggcc atggctttca tcctgtgtcc cctgacctgt ccaggtgagt gtgagggcag 2640 cactgggaag ctggagtgct gcttgtgcct cccttcccag tgggctgtgt tgactgctgc 2700 tccccacccc taccgatggt cccaggaagc agggagagtt ggggaaggca agattggaaa 2760 gacaggaaga ccaaggcctc ggcagaactc tctgtcttct ctccacttct ggtcccctgt 2820 ggtgatgtgc ctgtaatctt tttctccacc caaacccctt cccacgacaa aaacaagact 2880 gcctccctct cttccgggag ctggtgacag ccttgggcct ttcagtccca aagcggccga 2940 tgggagtctc cctccgactc cagatatgaa cagggcccag gcctggagcg tttgctgtgc 3000 caggaggcgg cagctcttct gggcagagcc tgtccccgcc ttccctcact cttcctcatc 3060 ctgcttctct tttcctcgca gatgataaaa ggaatctggc attctacacc tggaccattt 3120 gattgtttta ttttggaatt ggtgtatatc atgaagcctt gctgaactaa gttttgtgtg 3180 tatatattta aaaaaaaaat cagtgtttaa ataaagacct atgtacttaa tcctttaact 3240 ctgcggatag catttggtag gtagtgatta actgtgaata ataaatacac aatgaattct 3300 tcaaaaaaaa aaaaaaa 3317 <210> 114 <211> 4772 <212> DNA <213> human <400> 114 gctcagttgc cgggcggggg agggcgcgtc cggtttttct caggggacgt tgaaattatt 60 tttgtaacgg gagtcgggag aggacggggc gtgccccgac gtgcgcgcgc gtcgtcctcc 120 ccggcgctcc tccacagctc gctggctccc gccgcggaaa ggcgtcatgc cgcccaaaac 180 cccccgaaaa acggccgcca ccgccgccgc tgccgccgcg gaacccccgg caccgccgcc 240 gccgccccct cctgaggagg acccagagca ggacagcggc ccggaggacc tgcctctcgt 300 caggcttgag tttgaagaaa cagaagaacc tgattttact gcattatgtc agaaattaaa 360 gataccagat catgtcagag agagagcttg gttaacttgg gagaaagttt catctgtgga 420 tggagtattg ggaggttata ttcaaaagaa aaaggaactg tggggaatct gtatctttat 480 tgcagcagtt gacctagatg agatgtcgtt cacttttact gagctacaga aaaacataga 540 aatcagtgtc cataaattct ttaacttact aaaagaaatt gataccagta ccaaagttga 600 taatgctatg tcaagactgt tgaagaagta tgatgtattg tttgcactct tcagcaaatt 660 ggaaaggaca tgtgaactta tatatttgac acaacccagc agttcgatat ctactgaaat 720 aaattctgca ttggtgctaa aagtttcttg gatcacattt ttattagcta aaggggaagt 780 attacaaatg gaagatgatc tggtgatttc atttcagtta atgctatgtg tccttgacta 840 ttttattaaa ctctcacctc ccatgttgct caaagaacca tataaaacag ctgttatacc 900 cattaatggt tcacctcgaa cacccaggcg aggtcagaac aggagtgcac ggatagcaaa 960 acaactagaa aatgatacaa gaattattga agttctctgt aaagaacatg aatgtaatat 1020 agatgaggtg aaaaatgttt atttcaaaaa ttttatacct tttatgaatt ctcttggact 1080 tgtaacatct aatggacttc cagaggttga aaatctttct aaacgatacg aagaaattta 1140 tcttaaaaat aaagatctag atgcaagatt atttttggat catgataaaa ctcttcagac 1200 tgattctata gacagttttg aaacacagag aacaccacga aaaagtaacc ttgatgaaga 1260 ggtgaatgta attcctccac acactccagt taggactgtt atgaacacta tccaacaatt 1320 aatgatgatt ttaaattcag caagtgatca accttcagaa aatctgattt cctattttaa 1380 caactgcaca gtgaatccaa aagaaagtat actgaaaaga gtgaaggata taggatacat 1440 ctttaaagag aaatttgcta aagctgtggg acagggttgt gtcgaaattg gatcacagcg 1500 atacaaactt ggagttcgct tgtattaccg agtaatggaa tccatgctta aatcagaaga 1560 agaacgatta tccattcaaa attttagcaa acttctgaat gacaacattt ttcatatgtc 1620 tttattggcg tgcgctcttg aggttgtaat ggccacatat agcagaagta catctcagaa 1680 tcttgattct ggaacagatt tgtctttccc atggattctg aatgtgctta atttaaaagc 1740 ctttgatttt tacaaagtga tcgaaagttt tatcaaagca gaaggcaact tgacaagaga 1800 aatgataaaa catttagaac gatgtgaaca tcgaatcatg gaatcccttg catggctctc 1860 agattcacct ttatttgatc ttattaaaca atcaaaggac cgagaaggac caactgatca 1920 ccttgaatct gcttgtcctc ttaatcttcc tctccagaat aatcacactg cagcagatat 1980 gtatctttct cctgtaagat ctccaaagaa aaaaggttca actacgcgtg taaattctac 2040 tgcaaatgca gagacacaag caacctcagc cttccagacc cagaagccat tgaaatctac 2100 ctctctttca ctgttttata aaaaagtgta tcggctagcc tatctccggc taaatacact 2160 ttgtgaacgc cttctgtctg agcacccaga attagaacat atcatctgga cccttttcca 2220 gcacaccctg cagaatgagt atgaactcat gagagacagg catttggacc aaattatgat 2280 gtgttccatg tatggcatat gcaaagtgaa gaatatagac cttaaattca aaatcattgt 2340 aacagcatac aaggatcttc ctcatgctgt tcaggagaca ttcaaacgtg ttttgatcaa 2400 agaagaggag tatgattcta ttatagtatt ctataactcg gtcttcatgc agagactgaa 2460 aacaaatatt ttgcagtatg cttccaccag gccccctacc ttgtcaccaa tacctcacat 2520 tcctcgaagc ccttacaagt ttcctagttc acccttacgg attcctggag ggaacatcta 2580 tatttcaccc ctgaagagtc catataaaat ttcagaaggt ctgccaacac caacaaaaat 2640 gactccaaga tcaagaatct tagtatcaat tggtgaatca ttcgggactt ctgagaagtt 2700 ccagaaaata aatcagatgg tatgtaacag cgaccgtgtg ctcaaaagaa gtgctgaagg 2760 aagcaaccct cctaaaccac tgaaaaaact acgctttgat attgaaggat cagatgaagc 2820 agatggaagt aaacatctcc caggagagtc caaatttcag cagaaactgg cagaaatgac 2880 ttctactcga acacgaatgc aaaagcagaa aatgaatgat agcatggata cctcaaacaa 2940 ggaagagaaa tgaggatctc aggaccttgg tggacactgt gtacacctct ggattcattg 3000 tctctcacag atgtgactgt ataactttcc caggttctgt ttatggccac atttaatatc 3060 ttcagctctt tttgtggata taaaatgtgc agatgcaatt gtttgggtga ttcctaagcc 3120 acttgaaatg ttagtcattg ttatttatac aagattgaaa atcttgtgta aatcctgcca 3180 tttaaaaagt tgtagcagat tgtttcctct tccaaagtaa aattgctgtg ctttatggat 3240 agtaagaatg gccctagagt gggagtcctg ataacccagg cctgtctgac tactttgcct 3300 tcttttgtag catataggtg atgtttgctc ttgtttttat taatttatat gtatattttt 3360 ttaatttaac atgaacaccc ttagaaaatg tgtcctatct atcttccaaa tgcaatttga 3420 ttgactgccc attcaccaaa attatcctga actcttctgc aaaaatggat attattagaa 3480 attagaaaaa aattactaat tttacacatt agattttatt ttactattgg aatctgatat 3540 actgtgtgct tgttttataa aattttgctt ttaattaaat aaaagctgga agcaaagtat 3600 aaccatatga tactatcata ctactgaaac agatttcata cctcagaatg taaaagaact 3660 tactgattat tttcttcatc caacttatgt ttttaaatga ggattattga tagtactctt 3720 ggtttttata ccattcagat cactgaattt ataaagtacc catctagtac ttgaaaaagt 3780 aaagtgttct gccagatctt aggtatagag gaccctaaca cagtatatcc caagtgcact 3840 ttctaatgtt tctgggtcct gaagaattaa gatacaaatt aattttactc cataaacaga 3900 ctgttaatta taggagcctt aatttttttt tcatagagat ttgtctaatt gcatctcaaa 3960 attattctgc cctccttaat ttgggaaggt ttgtgttttc tctggaatgg tacatgtctt 4020 ccatgtatct tttgaactgg caattgtcta tttatctttt atttttttaa gtcagtatgg 4080 tctaacactg gcatgttcaa agccacatta tttctagtcc aaaattacaa gtaatcaagg 4140 gtcattatgg gttaggcatt aatgtttcta tctgattttg tgcaaaagct tcaaattaaa 4200 acagctgcat tagaaaaaga ggcgcttctc ccctccccta cacctaaagg tgtatttaaa 4260 ctatcttgtg tgattaactt atttagagat gctgtaactt aaaatagggg atatttaagg 4320 tagcttcagc tagcttttag gaaaatcact ttgtctaact cagaattatt tttaaaaaga 4380 aatctggtct tgttagaaaa caaaatttta ttttgtgctc atttaagttt caaacttact 4440 attttgacag ttattttgat aacaatgaca ctagaaaact tgactccatt tcatcattgt 4500 ttctgcatga atatcataca aatcagttag tttttaggtc aagggcttac tatttctggg 4560 tcttttgcta ctaagttcac attagaatta gtgccagaat tttaggaact tcagagatcg 4620 tgtattgaga tttcttaaat aatgcttcag atattattgc tttattgctt ttttgtattg 4680 gttaaaactg tacatttaaa attgctatgt tactattttc tacaattaat agtttgtcta 4740 ttttaaaata aattagttgt taagagtctt aa 4772 <210> 115 <211> 1260 <212> DNA <213> human <400> 115 aaattgagcc cgcagcctcc cgcttcgctc tctgctcctc ctgttcgaca gtcagccgca 60 tcttcttttg cgtcgccagc cgagccacat cgctcagaca ccatggggaa ggtgaaggtc 120 ggagtcaacg gatttggtcg tattgggcgc ctggtcacca gggctgcttt taactctggt 180 aaagtggata ttgttgccat caatgacccc ttcattgacc tcaactacat ggtttacatg 240 ttccaatatg attccaccca tggcaaattc catggcaccg tcaaggctga gaacgggaag 300 cttgtcatca atggaaatcc catcaccatc ttccaggagc gagatccctc caaaatcaag 360 tggggcgatg ctggcgctga gtacgtcgtg gagtccactg gcgtcttcac caccatggag 420 aaggctgggg ctcatttgca ggggggagcc aaaagggtca tcatctctgc cccctctgct 480 gatgccccca tgttcgtcat gggtgtgaac catgagaagt atgacaacag cctcaagatc 540 atcagcaatg cctcctgcac caccaactgc ttagcacccc tggccaaggt catccatgac 600 aactttggta tcgtggaagg actcatgacc acagtccatg ccatcactgc cacccagaag 660 actgtggatg gcccctccgg gaaactgtgg cgtgatggcc gcggggctct ccagaacatc 720 atccctgcct ctactggcgc tgccaaggct gtgggcaagg tcatccctga gctgaacggg 780 aagctcactg gcatggcctt ccgtgtcccc actgccaacg tgtcagtggt ggacctgacc 840 tgccgtctag aaaaacctgc caaatatgat gacatcaaga aggtggtgaa gcaggcgtcg 900 gagggccccc tcaagggcat cctgggctac actgagcacc aggtggtctc ctctgacttc 960 aacagcgaca cccactcctc cacctttgac gctggggctg gcattgccct caacgaccac 1020 tttgtcaagc tcatttcctg gtatgacaac gaatttggct acagcaacag ggtggtggac 1080 ctcatggccc acatggcctc caaggagtaa gacccctgga ccaccagccc cagcaagagc 1140 acaagaggaa gagagagacc ctcactgctg gggagtccct gccacactca gtcccccacc 1200 acactgaatc tcccctcctc acagttgcca tgtagacccc ttgaagaggg gaggggccta 1260 <210> 116 <211> 974 <212> DNA <213> human <400> 116 gtttgcttag gtgaggcgca gacggggaag cggagccaac atgccagtgg cccggagctg 60 ggtttgtcgc aaaacttatg tgaccccgcg gagacccttc gagaaatctc gtctcgacca 120 agagctgaag ctgatcggcg agtatgggct ccggaacaaa cgtgaggtct ggagggtcaa 180 atttaccctg gccaagatcc gcaaggccgc ccgggaactg ctgacgcttg atgagaagga 240 cccacggcgt ctgttcgaag gcaacgccct gctgcggcgg ctggtccgca ttggggtgct 300 ggatgagggc aagatgaagc tggattacat cctgggcctg aagatagagg atttcttaga 360 gagacgcctg cagacccagg tcttcaagct gggcttggcc aagtccatcc accacgctcg 420 cgtgctgatc cgccagcgcc atatcaggta ccacctcgga tgggcacctg aatcttcctc 480 cacctgcccc tctgatggtt gccctcacta agcctgctgt ccctatctcc tatgcagccc 540 tcggaggtga tgggtgtgaa ctcacccaga gggtacagat tcacccttgc acacagctca 600 ccagggagct ggggcagcct cttgccccaa tagcccagcg caagggtcac tgcggctcta 660 gccgtacacc ttgtgaaggc ctctgccagg catgtgggca gctggacagg gtccgcaagc 720 aggtggtgaa catcccgtcc ttcattgtcc gcctggattc ccagaagcac atcgacttct 780 ctctgcgctc tccctacggg ggtggccgcc cgggccgcgt gaagaggaag aatgccaaga 840 agggccaggg tggggctggg gctggagacg acgaggagga ggattaagtc cacctgtccc 900 tcctgggctg ctggattgtc tcgttttcct gccaaataaa caggatcagc gctttaaaaa 960 aaaaaaaaaa aaaa 974

Claims (16)

A method of measuring the level of the gene marker Pim-1 to predict or monitor the response of cancer patients to paclitaxel, docetaxel, XRP9881 or XRP6258,
a) measuring the level of Pim-1 in test samples and control samples obtained from cancer patients; And
b) comparing the measured levels of Pim-1 in the test sample and the control sample
Wherein the decrease in the level of Pim-1 measured in the test sample compared to the control sample is indicative of increased sensitivity to paclitaxel, docetaxel, XRP9881 or XRP6258. The method of claim 1, wherein the level of Pim-1 is measured by mRNA, DNA or protein. The method of claim 2, wherein the mRNA is measured by in situ hybridization, reverse transcriptase polymerase chain reaction, nucleic acid hybridization, electrophoresis, northern blotting or mass spectrometry. The method of claim 2, wherein the DNA is measured by quantitative polymerase chain reaction, genomic DNA-chip, in situ hybridization, electrophoresis, southern blotting or mass spectrometry. The method of claim 2, wherein the protein is measured by immunological assay, western blot, ELISA or mass spectrometry. A kit for measuring the level of the gene marker Pim-1 to predict or monitor the response of a cancer patient to paclitaxel, docetaxel, XRP9881 or XRP6258 according to the method of claim 1,
A kit comprising a detectable-labeled antibody, a detectable-labeled antibody fragment or a detectable-labeled oligonucleotide, comprising a nucleic acid capable of hybridizing to Pim-1 under stringent conditions. A kit for measuring the level of the gene marker Pim-1 to predict or monitor the response of a cancer patient to paclitaxel, docetaxel, XRP9881 or XRP6258 according to the method of claim 1,
a) means for obtaining total RNA from a sample of body origin;
b) means for reverse transcription of the total RNA to obtain cDNA; And
c) means for applying said cDNA to a polymerase chain reaction using a set of primers wherein one or both primers are detectably-labeled and both primers are derived from the nucleotide sequence of Pim-1
&Lt; / RTI &gt; 8. The kit of claim 6 or 7, wherein the detectable label is an enzyme, radioisotope, fluorescent chemical, chemiluminescent molecule, radiopaque material, liposome, or hapten molecule. A method of measuring the level of the gene marker Pim-1 to predict or monitor the response of cancer patients to paclitaxel, docetaxel, XRP9881 or XRP6258,
a) measuring the level of Pim-1 in a test sample obtained from the cancer area of a cancer patient;
b) measuring the level of one or more reference genetic markers; And
c) comparing the measured levels of Pim-1 and said at least one reference genetic marker in a test sample
Wherein the decrease in the level of Pim-1 in comparison to the level of the one or more reference genetic markers indicates increased sensitivity to paclitaxel, docetaxel, XRP9881 or XRP6258. The method of claim 9, wherein the level of Pim-1 is measured by mRNA, DNA or protein. The method of claim 10, wherein the mRNA is measured by in situ hybridization, reverse transcriptase polymerase chain reaction, nucleic acid hybridization, electrophoresis, northern blotting, or mass spectrometry. The method of claim 10, wherein the DNA is measured by quantitative polymerase chain reaction, genomic DNA-chip, in situ hybridization, electrophoresis, southern blotting or mass spectrometry. The method of claim 10, wherein the protein is measured by immunological assay, western blot, ELISA or mass spectrometry. A kit for measuring the level of the gene marker Pim-1 to predict or monitor the response of a cancer patient to paclitaxel, docetaxel, XRP9881 or XRP6258 according to the method of claim 9,
A kit comprising a detectable-labeled antibody, a detectable-labeled antibody fragment or a detectable-labeled oligonucleotide, comprising a nucleic acid capable of hybridizing to Pim-1 under stringent conditions. A kit for measuring the level of the gene marker Pim-1 to predict or monitor the response of a cancer patient to paclitaxel, docetaxel, XRP9881 or XRP6258 according to the method of claim 9,
a) means for obtaining total RNA from a sample of body origin;
b) means for reverse transcription of the total RNA to obtain cDNA; And
c) means for applying said cDNA to a polymerase chain reaction using a set of primers wherein one or both primers are detectably-labeled and both primers are derived from the nucleotide sequence of Pim-1
&Lt; / RTI &gt; The kit of claim 14 or 15, wherein the detectable label comprises an enzyme, radioisotope, fluorescent chemical, chemiluminescent molecule, radiopaque material, liposome, or hapten molecule.

Images